Biofilm Layer Research Articles

PDA-Fe3O4 decorated carbon felt anode enhancing electrochemical performance of microbial fuel cells: Effect of electrode materials on electroactive biofilm

Anode modification is an effective strategy for enhancing the electrochemical performance of microbial fuel cell (MFC). However, the impacts of the modified materials on anode biofilm development during MFC operation have been less studied. We prepared a novel PDA-Fe3O4-CF composite anode by coating original carbon felt anode (CF) with polydopamine (PDA) and Fe3O4 nanoparticles. The composite anode material was characterized by excellent hydrophilicity and electrical conductivity, and the anodic biofilm exhibited fast start-up, higher biomass, and more uniform biofilm layer after MFC operation. The MFC reactor assembled with the composite anode achieved a maximum power density of 608 mW m−2 and an output voltage of 586 mV, which were 316.4% and 72.4% higher than the MFC with the original CF anode, respectively. Microbial community analysis indicated that the modified anode biofilm had a higher relative abundance of exoelectrogen species in comparison to the unmodified anode. The PICRUSt data revealed that the anodic materials may affect the bioelectrochemical performance of the biofilm by influencing the expression levels of key enzyme genes involved in biofilm extracellular polymer (EPS) secretion and extracellular electron transfer (EET). The growth of the anodic biofilm would exert positive or negative influences on the efficiency of electricity production and electron transfer of the MFCs at different operating stages. This work expands the knowledge of the role that anodic materials play in the development and electrochemical performance of anodic biofilm in MFCs.

Bioaccumulation of trace metals in the plastisphere: Awareness of environmental risk from a European perspective

The term "Plastisphere" refers to the biofilm layer naturally formed by microorganisms attaching to plastic surfaces. This layer possesses the capability to adsorb persistent organic and inorganic pollutants, particularly trace metals, which are the focus of this research study. Immersion experiments were concurrently conducted in five locations spanning four European countries (France, Ireland, Spain, and Italy) utilising eight distinct polymers. These immersions, repeated every three months over a one-year period, aimed to evaluate the baseline bioaccumulation of 12 trace metals. The study underscores the intricate nature of metal bioaccumulation, influenced by both micro-scale factors (such as polymer composition) and macro-scale factors (including geographical site and seasonal variations). Villefranche Bay in France exhibited the lowest metals bioaccumulation, whereas Naples in Italy emerged as the site where bioaccumulation was often the highest for the considered metals. Environmental risk assessment was also conducted in the study.The lightweight nature of certain plastics allows them to be transported across significant distances in the ocean. Consequently, evaluating trace metal concentrations in the plastisphere is imperative for assessing potential environmental repercussions that plastics, along with their associated biota, may exert even in locations distant from their point of emission.

Sub-inhibitory concentrations of colistin and imipenem impact the expression of biofilm-associated genes in Acinetobacter baumannii.

Acinetobacter baumannii is an opportunistic pathogen that is responsible for nosocomial infections. Imipenem and colistin are drugs that are commonly used to treat severe infections caused by A. baumannii, such as sepsis, ventilator-associated pneumonia, and bacteremia. However, some strains of A. baumannii have become resistant to these drugs, which is a concern for public health. Biofilms produced by A. baumannii increase their resistance to antibiotics and the cells within the inner layers of biofilm are exposed to sub-inhibitory concentrations (sub-MICs) of antibiotics. There is limited information available regarding how the genes of A. baumannii are linked to biofilm formation when the bacteria are exposed to sub-MICs of imipenem and colistin. Thus, this study's objective was to explore this relationship by examining the genes involved in biofilm formation in A. baumannii when exposed to low levels of imipenem and colistin. The study found that exposing an isolate of A. baumannii to low levels of these drugs caused changes in their drug susceptibility pattern. The relative gene expression profiles of the biofilm-associated genes exhibited a change in their expression profile during short-term and long-term exposure. This study highlights the potential consequences of overuse and misuse of antibiotics, which can help bacteria become resistant to these drugs.

Biofilm formation monitoring using SEM in synthetic wastewater pollutant removal by combination of DAF and modified MBBR

Nowadays, moving bed biofilm reactors (MBBR) and dissolved air flotation (DAF) are frequently used in combination for the biological and physical treatments of synthetic oily wastes. The efficiency of the biocarriers utilized in MBBR is crucial since it has a big impact on the system's overall performance. A study was carried out utilizing a DAF-MBBR technique to treat synthetic oily wastewater in order to evaluate this. Two different kinds of biocarriers were used in this modified MBBR: polyethylene and plastic bottle caps. The result showed the DAF-MMBBRs process demonstrated excellent removal efficiencies for a range of parameters. Moreover, it seemed that DAF had higher COD, nitrite, nitrate, and turbidity removal efficiencies. For COD, the average removal efficiency was 39.62%; for oil and grease, it was 77.05%; and for ammonia, it was 39.71%. In the same way, MBBR1 is effective at eliminating phosphate, ammonia, TSS, and oil and grease. For COD, the average removal efficiency was 39.12%; for oil and grease, it was 94.10%; and for ammonia, it was 85.42%. The average removal efficiencies for COD, oil and grease, and ammonia in MMBBR2 were 38.76%, 84.95%, and 86.49%, respectively. Furthermore, Scanning electron microscopy (SEM) was used to assess the biofilm within the system. This revealed a well-developed biofilm layer on the interior surfaces of the biocarriers, improving the effectiveness of synthetic wastewater treatment. Excellent removal efficiencies were shown by the DAF-MBBR system for a range of parameters during a ten-day operational period.

Enterococcus mundtii A2 biofilm and its anti-adherence potential against pathogenic microorganisms on stainless steel 316L.

Pathogenic bacterial biofilms present significant challenges, particularly in food safety and material deterioration. Therefore, using Enterococcus mundtii A2, known for its antagonistic activity against pathogen adhesion, could serve as a novel strategy to reduce pathogenic colonization within the food sector. This study aimed to investigate the biofilm-forming ability of E. mundtii A2, isolated from camel milk, on two widely used stainless steels within the agri-food domain and to assess its anti-adhesive properties against various pathogens, especially on stainless steel 316L. Additionally, investigations into auto-aggregation and co-aggregation were also conducted. Plate count methodologies revealed increased biofilm formation by E. mundtii A2 on 316L, followed by 304L. Scanning electron microscopy (SEM) analysis revealed a dense yet thin biofilm layer, playing a critical role in reducing the adhesion of L. monocytogenes CECT 4032 and Staphylococcus aureus CECT 976, with a significant reduction of ≈ 2 Log CFU/cm2. However, Gram-negative strains, P. aeruginosa ATCC 27853 and E. coli ATCC 25922, exhibit modest adhesion reduction (~ 0.7 Log CFU/cm2). The findings demonstrate the potential of applying E. mundtii A2 biofilms as an effective strategy to reduce the adhesion and propagation of potentially pathogenic bacterial species on stainless steel 316L.

Improving Activity of New Arylurea Agents against Multidrug-Resistant and Biofilm-Producing Staphylococcus epidermidis.

Multidrug-resistant (MDR) strains of Staphylococcus epidermidis (S. epidermidis), prevalent in hospital environments, contribute to increased morbidity and mortality, especially among newborns, posing a critical concern for neonatal sepsis. In response to the pressing demand for novel antibacterial therapies, we present findings from synthetic chemistry and structure-activity relationship studies focused on arylsulfonamide/arylurea derivatives of aryloxy[1-(thien-2-yl)propyl]piperidines. Through bioisosteric replacement of the sulfonamide fragment with a urea moiety, compound 25 was identified, demonstrating potent bacteriostatic activity against clinical multidrug-resistant S. epidermidis strains (MIC50 and MIC90 = 1.6 and 3.125 μg/mL). Importantly, it showed activity against linezolid-resistant strains and exhibited selectivity over mammalian cells. Compound 25 displayed antibiofilm-forming properties against clinical S. epidermidis strains and demonstrated the capacity to eliminate existing biofilm layers. Additionally, it induced complete depolarization of the bacterial membrane in clinical S. epidermidis strains. In light of these findings, targeting bacterial cell membranes with compound 25 emerges as a promising strategy in the fight against multidrug-resistant S. epidermidis strains.

Chitosan modified magnetic nanocomposite for biofilm destruction and precise photothermal/photodynamic therapy

Getting rid of the biofilms is a major challenge when treating skin and soft tissue infections (SSTI), an inflammatory illness brought on by bacteria. Traditional magnetic materials have a limited dispersibility and a biofilm permeable property, making it challenging to remove biofilms and causing infection to linger. To solve these problems, we developed a kind of magnetic composite nanoplatform coated with indocyanine green carbon dots and modified with chitosan modification (Fe-ICGCDs@CS). Fe-ICGCDs@CS has high dispersibility and improves the conductivity of biofilms under magnetic action. Fe-ICGCDs@CS can adsorb bacteria via the positive charge and achieve precise photothermal sterilization and photodynamic therapy (PDT). Moreover, by catalyzing hydrogen peroxide (2 mM), Fe-ICGCDs@CS can produce oxygen to relieve the anoxic state in the deep layer of biofilms and activate dormant bacteria to make them sensitive to external stimuli. All in all, unlike the common “just kill” sterilization model, Fe-ICGCDs@CS can accurately kill bacteria and be recovered by an external magnetic field at the end of treatment, thus reducing the potential biological toxicity of nanomaterials. Therefore, the proposed Fe-ICGCDs@CS provides a new antibacterial method with low biotoxicity for clinical application in the treatment of biofilm infections.

Characterization, antibacterial, and cytotoxic activities of silver nanoparticles using the whole biofilm layer as a macromolecule in biosynthesis

Recently, multi-drug resistant (MDR) bacteria are responsible for a large number of infectious diseases that can be life-threatening. Globally, new approaches are targeted to solve this essential issue. This study aims to discover novel antibiotic alternatives by using the whole components of the biofilm layer as a macromolecule to synthesize silver nanoparticles (AgNPs) as a promising agent against MDR. In particular, the biosynthesized biofilm-AgNPs were characterized using UV-Vis spectroscopy, electron microscopes, Energy Dispersive X-ray (EDX), zeta sizer and potential while their effect on bacterial strains and normal cell lines was identified. Accordingly, biofilm-AgNPs have a lavender-colored solution, spherical shape, with a size range of 20–60 nm. Notably, they have inhibitory effects when used on various bacterial strains with concentrations ranging between 12.5 and 25 µg/mL. In addition, they have an effective synergistic effect when combined with phage ZCSE9 to inhibit and kill Salmonella enterica with a concentration of 3.1 µg/mL. In conclusion, this work presents a novel biosynthesis preparation of AgNPs using biofilm for antibacterial purposes to reduce the possible toxicity by reducing the MICs using phage ZCSE9.

Long-term Evaluation of Culturable Fungi in a Natural Aging Biofilm on Galvanised Steel Surface

Fungi commonly found in municipal water can participate in natural biofilm formation on the surface of galvanised steel despite the toxic effect of zinc. Depending on the age of the biofilm, fungal diversity may vary. To examine this hypothesis, natural biofilm formation was allowed on galvanised steel surfaces over six months in a model recirculating water system. Fungal colonies with different morphologies were obtained monthly from biofilm and water samples and then identified by both morphological and molecular approaches. In addition, the biofilm layer was examined by electrochemical impedance spectroscopy (EIS) analysis and scanning electron microscopy (SEM). It was determined that fungi were included in the naturally aging biofilm formed on galvanised steel surfaces during the experiment. The diversity and the number of fungi in the biofilm and water changed over the experiment. All fungi isolated from the biofilm and water were found to be members of the Ascomycota phylum. F. oxysporum was the first fungus to be involved in the biofilm formation process and was one of the main inhabitants of the biofilm together with Penicillium spp. In addition, EIS data showed that the structure of the biofilm changed as it aged. The results of this study may lead to a better understanding of naturally aging biofilms involving fungi in municipal water systems, as well as the development of new strategies for effective disinfection of fungi based on biofilm age.

Effect of N-acetyl cysteine, rifampicin, and ozone on biofilm formation in pan-resistant Klebsiella pneumoniae: an experimental study.

To the best of our knowledge, this is the first study to evaluate the effectiveness of specific concentrations of antibiofilm agents, such as N-acetyl cysteine (NAC), rifampicin, and ozone, for the treatment of pan-resistant Klebsiella pneumoniae (PRKp). We evaluated the effectiveness of antibiofilm agents, such as NAC, rifampicin, and ozone, on biofilm formation in PRKp at 2, 6, 24, and 72 h. This single-center experimental study was conducted on June 15, 2017, and July 15, 2018, at Istanbul Faculty of Medicine, Istanbul University, Turkey. Biofilm formation and the efficacy of these agents on the biofilm layer were demonstrated using colony counting and laser-screened confocal microscopy. NAC at a final concentration of 2 μg/mL was administered to bacteria that formed biofilms (24 h), and no significant decrease was detected in the bacterial counts of all isolates (all P > 0.05). Rifampicin with a final concentration of 0.1 μg/mL was administered to bacteria that formed biofilm (24 h), and no significant decrease was detected in bacterial count (all P > 0.05). Notably, ozonated water of even 4.78 mg/L concentration for 72 h decreased the bacterial count by ≥ 2 log10. Different approaches are needed for treating PRKp isolates. We demonstrate that PRKp isolates can be successfully treated with higher concentrations of ozone.

Self-controllable proteinic antibacterial coating with bacteria-triggered antibiotic release for prevention of periprosthetic infection

Periprosthetic infection is a devastating postimplantation complication in which a biofilm layer harboring invasive microorganisms forms around orthopedic implants, leading to severe implant failure and patient morbidity. Despite the development of several infection-triggered antibiotic release approaches, most current antibacterial coatings are susceptible to undesired antibiotic leakage or mechanical disintegration during prosthesis installation. Herein, we propose a self-controllable proteinic antibacterial coating capable of both long-lasting adherence onto titanium implant substrates over the implant fixation period and instantaneous bacterial eradication. Importantly, the pH-dependent reversible metal coordination of mussel adhesive protein (MAP) enabled bacterial concentration-dependent antibiotic delivery in response to infection-induced acidification. In addition, the MAP coating exhibited superior self-healable adhesive properties and scratch resistance, which enabled to avert issues associated with mechanical damages, including peeling and cracking, often occurring in conventional implant coating systems. The gentamicin-loaded MAP coating exhibited complete inhibition of bacterial growth in vivo against Staphylococcus aureus penetrations during implantation surgery (immediate infection) and even 4 weeks after implantation (delayed infection). Thus, our antibiotic-loaded MAP hydrogel coating can open new avenues for self-defensive antibiotic prophylaxis to achieve instant and sustainable bacteriocidal activity in orthopedic prostheses.© 2017 Elsevier Inc. All rights reserved.

Features of the rare pathogen Meyerozyma guilliermondii strain SO and comprehensive in silico analyses of its adherence-contributing virulence factor agglutinin-like sequences

Meyerozyma guilliermondii is a rare yeast pathogen contributing to the deadly invasive candidiasis. M. guilliermondii strain SO, as a promising protein expression host, showed 99% proteome similarity with the clinically isolated ATCC 6260 (type strain) in a recent comparative genomic analysis. However, their in vitro virulence features and in vivo pathogenicity were uncharacterized. This study aimed to characterize the in vitro and in vivo pathogenicity of M. guilliermondii strain SO and analyze its Als proteins (MgAls) via comprehensive bioinformatics approaches. M. guilliermondii strain SO showed lower and higher sensitivity towards β-mercaptoethanol and lithium, respectively than the avirulent S. cerevisiae but exhibited the same tolerance towards cell wall-perturbing Congo Red with C. albicans. With 7.5× higher biofilm mass, M. guilliermondii strain SO also demonstrated 75% higher mortality rate in the zebrafish embryos with a thicker biofilm layer on the chorion compared to the avirulent S. cerevisiae. Being one of the most important Candida adhesins, sequence and structural analyses of four statistically identified MgAls showed that MgAls1056 was predicted to exhibit the most conserved amyloid-forming regions, tandem repeat domain and peptide binding cavity (PBC) compared to C. albicans Als3. Favoured from the predicted largest ligand binding site and druggable pockets, it showed the highest affinity towards hepta-threonine. Non-PBC druggable pockets in the most potent virulence contributing MgAls1056 provide new insights into developing antifungal drugs targeting non-albicans Candida spp. Virtual screening of available synthetic or natural bioactive compounds and MgAls1056 deletion from the fungal genome should be further performed and validated experimentally. Communicated by Ramaswamy H. Sarma

(Invited) Stabilization and Activation of Copper(I)-Oxide-Semiconducting Interfaces for Photoelectrochemical Reduction of Carbon Dioxide

Electroreduction of carbon dioxide to simple organic fuels and chemicals is a topic of growing scientific and technological interest. The reaction provides means for both reducing emissions of CO2 into atmosphere and storing renewable energy. The presentation will address low-temperature CO2-conversion processes based on electrocatalytic and photoelectrochemical approaches. Among important issues are choice of the catalytic or semiconducting materials, their morphology and operating conditions including temperature, solvent, electrolyte, pH etc. There is a need to improve the reaction dynamics and selectivity toward specific products. In practical electrolysis cells, the CO2-reduction (at cathode) is accompanied by water oxidation (at anode or photoanode).Recently, we have concentrated on the development of hybrid materials by utilizing combination of metal oxide semiconductors thus capable of effective photoelectrochemical reduction of carbon dioxide. For example, the combination of conducting polymers, or titanium (IV) oxide, and copper (I) oxide has been considered before and after sunlight illumination. Application of the hybrid system composed of both above-mentioned oxides resulted in high current densities originating from photoelectrochemical reduction of carbon dioxide mostly to methanol (CH3OH) as demonstrated upon identification of final products. Among important issue is intentional stabilization, activation, and functionalization of the mixed-metal-oxide-based photoelectrochemcal interface toward better long-term performance and selectivity production of small organic molecules (C1-C4) and other chemicals. In this respect, ultra-thin films of conducting polymers (simple or polyoxometallate-derivatized) and supramolecular complexes (with nitrogen containing ligands and certain transition metal sites), sub-monolayers of metals (Cu, Au), networks of noble metal (Au, Ag) nanoparticles or layers of robust bacterial biofilms have been considered. The photobiocathode utilizing robust biofilms have also been demonstrated to stabilize copper(I) oxide surfaces and to induce the system’s activity toward reduction of carbon dioxide under illuminations with visible light.In the presentation, special attention will be paid to the mechanistic aspects of electroreduction of carbon dioxide, fabrication and characterization of highly selective and durable semiconductor photoelectrode materials and to the importance of the reaction conditions.

Relation Of Habitual Of Brushing Teeth With Candida sp. Colony Count in Children Dental Caries Under 10 Years Old

The dental and oral disease problem faced by the Indonesian population as well as in other developing countries is dental caries tissue disease. Bacteria colonize the oral cavity to produce acid, resulting in a demineralization process in dental caries tissue. Candida albicans is the main agent of oral candidiasis. The development of these two microorganisms interacts with each other to form a layer of biofilm, which is the beginning of caries. Candidiasis is a fungal infection caused by Candida sp., which is the normal microflora in the oral cavity. This study aims to analyze the relationship between tooth brushing habits and the number of Candida sp. in dental caries in children under 10 years of age. This research is an analytic survey with a cross-sectional approach. Based on research on the relationship between brushing teeth and the number of Candida sp. in children under 10 years of age at SDN 01 Petung Bondowoso students, it can be seen that out of a total of 13 respondents (100%) who have poor tooth brushing habits, there are 7 respondents (53.8%) who have a small number of colonies and 6 respondents (46.2%) who have many colonies. Meanwhile, out of a total of 41 respondents (100%) who had the habit of brushing their teeth well, it was found that 41 respondents (100%) had a small number of colonies and none had many colonies. Based on the Chi-Square test, the results obtained were p = 0.000. Because p = 0.000 0.05, it can be interpreted that there is a relationship between brushing teeth and the number of candida fungal colonies in dental caries in children under 10 years of age. This can also be interpreted as meaning that the better the habit of brushing your teeth, the fewer the number of candida fungal colonies, and vice versa.

Enhance the resistance of probiotics by microencapsulation and biofilm construction based on rhamnogalacturonan I rich pectin

Microcapsules were always used as functional material carriers for targeted delivery and meanwhile offering protection. However, microcapsule wall materials with specific properties were required, which makes the choice of wall material a key factor. In our previous study, a highly branched rhamnogalacturonan I rich (RG-I-rich) pectin was extracted from citrus canning processing water, which showed good gelling properties and binding ability, indicating it could be a potential microcapsule wall material. In the present study, Lactiplantibacillus plantarum GDMCC 1.140 and Lactobacillus rhamnosus were encapsulated by RG-I-rich pectin with embedding efficiencies of about 65 %. The environmental tolerance effect was evaluated under four different environmental stresses. Positive protection results were obtained under all four conditions, especially under H2O2 stress, the survival rate of probiotics embedded in microcapsules was about double that of free probiotics. The storage test showed that the total plate count of L. rhamnosus encapsulated in RG-I-rich pectin microcapsules could still reach 6.38 Log (CFU/mL) at 25 °C for 45 days. Moreover, probiotics embedded in microcapsules with additional incubation to form a biofilm layer inside could further improve the probiotics' activities significantly in the above experiments. In conclusion, RG-I-rich pectin may be a good microcapsule wall material for probiotics protection.

Antibiofilm activity of biosynthesized silver and copper nanoparticles using Streptomyces S29

Microbial resistance and biofilm formation have been considered as the main problems associated with microbial resistance. Several antimicrobial agents cannot penetrate biofilm layers and cannot eradicate microbial infection. Therefore, the aim of this study is the biological synthesis of silver and copper nanoparticles to assess their activities on bacterial attachment and on the viability of dormant cells within the biofilm matrix. Ag-NPs and Cu-NPs were biosynthesized using Streptomyces isolate S29. The biologically synthesized Ag-NPs and Cu-NPs exhibited brown and blue colors and were detected by UV/Vis spectrophotometry at 476 and 594 nm, respectively. The Ag-NPs showed an average size of 10–20 nm as indicated by TEM, and 25–35 nm for Cu-NPs. Both Ag-NPs and Cu-NPs were monodispersed with a polydispersity index of 0.1–0.546 and zeta potential were − 29.7, and − 33.7 mv, respectively. The biologically synthesized Ag-NPs and Cu-NPs significantly eliminated bacterial attachment and decreased the viable cells in the biofilm matrix as detected by using crystal violet and tri-phenyl tetrazolium chloride assays. Furthermore, Ag-NPs and Cu-NPs significantly eradicated mature biofilms developed by various Gram-negative pathogens, including A. baumannii, K. pneumoniae and P. aeruginosa standard strains and clinical isolates. Data were also confirmed at the molecular level with prominent elimination of biofilm gene expression carO, bssS and pelA in A. baumannii, K. pneumoniae and P. aeruginosa, respectively compared to untreated cells under the same conditions. As indicated, Ag-NPs and Cu-NPs could be used as adjuvant therapy in eradication of antibiotic resistance and biofilm matrix associated with Gram-negative bacterial infection.Graphical

Optimizing cleaning strategies for biofouling in reverse osmosis membrane systems: A comparative study using a self-formed instrument

Membrane biofouling poses a serious challenge in reverse osmosis (RO) plants used for water recovery, which cannot be addressed by traditional chlorine-based cleaning agents. To accommodate this problem and enhance membrane performance while maintaining high-quality water production, various cleaning strategies have been applied. It remains however challenging to identify optimal cleaning agents, as tests require discontinuation of production to evaluate the effect. This study introduces a self-formed instrument that mimics industrial RO plants on a small scale to compare the cleaning agents in a fast, affordable and feasible approach. The setup was demonstrated using three cells operating in parallel and a thorough comparison of these. Specifically, the online cleaning efficiency of a typical biofouling cleaner, a solution of NaOH/sodium dodecyl sulfate (SDS), and the commonly used non-oxidizing biocide, 2,2-dibromo-3-nitrilopropionamide, were compared to validate the system performance when fed with membrane bioreactor effluent. The effectiveness of the cleaning process was evaluated using a combination of hydrodynamic operation and membrane autopsy to estimate the biofoulant amount and composition. During a 7-day operating period, it was observed that treated membranes demonstrated higher flux rates compared to untreated membranes. The normalized flux increased more than double in the membrane after it had been cleaned with NaOH/SDS solution. CLSM analysis clearly showed the distinct effects of the two cleaning solutions on the amount and spatial structure of the biofilm layer. Cleaning with DBNPA was able to reduce the load of living bacteria, but not necessarily remove the biofilm components completely. Comparative analyses of membranes revealed that NaOH/SDS exhibited the highest cleaning efficiency during the online cleaning operation mode. This benchmark highlights the versatility of the system, which can be assembled at relevant sites and assist in the identification of efficient and eco-friendly agents that promote the lifespan of membranes. With the distribution of this instrument, it will be possible to accelerate the investigation of multiple cleaning strategies and also analyze interactions among cleaning agents in an attempt to reduce water shortage.

Effect of N/P ratio on attached microalgae growth and the differentiated metabolism along the depth of biofilm

Attached microalgae cultivation coupled with wastewater treatment could convert pollutants into bioresource with high efficiency and low cost. Nitrogen to phosphorus ratio (N/P ratio) is considered as an important factor on microalgae growth. Due to spatially heterogeneous distribution of nutrient, how N/P ratio affected attached microalgae growth in both macro- and micro-scopes was explored in this study. The findings revealed that an optimal N/P ratio of 10:1 promoted attached microalgae growth, while unsuitable ratios hampered algal growth by inhibiting photosynthesis, lowering oxidative resistance and decreasing metabolism activity. Long-term cultivation with improper N/P ratios resulted in a gradual decrease in actual photosynthetic rates, implying 50 days as the upper culture time limit for high-efficiency growth. Moreover, the study highlighted the uneven distribution of light and nutrients in algal biofilms, causing cells in different biofilm layers with variability of metabolism and composition. However, the 15N isotopic distribution demonstrated that even bottom cells were equally capable of nitrogen assimilation.

Challenges of suppressing nitrite-oxidizing bacteria in membrane aerated biofilm reactors by low dissolved oxygen control

Membrane aerated biofilm reactor (MABR) and shortcut nitrogen removal are two types of solutions to reduce energy consumption in wastewater treatment, with the former improving the aeration efficiency and the latter reducing the oxygen demand. However, integrating these two solutions, i.e., achieving shortcut nitrogen removal in MABR, is challenging due to the difficulty in suppressing nitrite-oxidizing bacteria (NOB). In this study, four MABRs were established to demonstrate the feasibility of initiating, maintaining, and restoring NOB suppression using low dissolved oxygen (DO) control, in the presence and absence of anammox bacteria, respectively. Long-term results revealed that the strict low DO (< 0.1 mg/L) in MABR could initiate and maintain stable NOB suppression for more than five months with nitrite accumulation ratio above 90 %, but it was unable to re-suppress NOB once they prevailed. Moreover, the presence of anammox bacteria increased the threshold of DO level to maintain NOB suppression in MABRs, but it was still incapable to restore the deteriorated NOB suppression in conjunction with low DO control. Mathematical modelling confirmed the experimental results and further explored the differences of NOB suppression in conventional biofilms and MABR biofilms. Simulation results showed that it is more challenging to maintain stable NOB suppression in MABRs compared to conventional biofilms, regardless of biofilm thickness or influent nitrogen concentration. Kinetic mechanisms for NOB suppression in different types of biofilms were proposed, suggesting that it is difficult to wash out NOB developed in the innermost layer of MABR biofilms because of the high oxygen level and low sludge wasting rate. In summary, this study systematically demonstrated the challenges of NOB suppression in MABRs through both experiments and mathematical modelling. These findings provide valuable insights into the applications of MABRs and call for more studies in developing effective strategies to achieve stable shortcut nitrogen removal in this energy-efficient configuration.

Marine antifouling properties of enzyme modified polyaniline coated stainless steel surface

On solid surfaces immersed in a liquid medium, a biofilm layer which is called biofouling formed over time by organic molecules and microorganisms. It is important to produce new eco-friendly ideas can prevent this undesired phenome. In this study, we focused on the antifouling performance of polyaniline (PANI), whose anticorrosive properties have been already known. The main purpose of this study was to immobilize hydrolytic enzymes that could break down biomolecules and microorganisms and how this would contribute to the antifouling performance of the PANI coating. When α-amylase, DNAse, glucose oxidase, α-chymotrypsin, lipase and pectinase enzymes immobilized into PANI that was synthesized in ammonium oxalate (PANIAO) and sodium salicylate (PANISS) electrolytes, α-amylase containing film (PANISS-A) showed the highest performance (76.5% antifouling activity). The surface properties after keeping in the Mediterranean Sea for 12 days were compared by digital photography, Scanning Electron microscope (SEM) and fluorescence microscope images, also with Energy Dispersive X-Ray (EDX) analysis and crystal violet staining. Carbohydrate and protein amounts and CFU (Colony Forming Units) values of biofilms formed on the surface for bare, PANISS and PANISS-A coupons after keeping 12 days in the Mediterranean Sea were determined. Vibrio species (V.harveyi, V.alginolyticus, V.parahaemolyticus) were detected in the biofilms by Matrix- Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) analysis.