Biofilm Age Research Articles

Evaluation of Salmonella biofilm attachment and hydrophobicity characteristics on food contact surfaces

Salmonella forms biofilms, and persist on food contact surfaces. Once a biofilm is formed cleaning and sanitation protocols may be inadequate for effective removal. This study evaluated attachment characteristics, surface properties, and structure of Salmonella biofilms on food contact surfaces commonly used in the tree-fruit industry. Multi-strain Salmonella biofilms were grown in a Centers for Disease Control and Prevention (CDC) biofilm reactor at 22 ± 2 °C and sampling was conducted at 2, 24 and 96-h. After each incubation period, coupons weregently rinsed and the remaining cells enumerated. Biofilms were analyzed with Laser Scanning Confocal Microscopy (LSCM). Hydrophobicity was evaluated by measuring the contact angles of reference liquids method using a drop tensiometer instrument. Material type and biofilm age significantly influenced attachment and biofilm hydrophobicity (P < 0.05). The strength of attachment, across all time points, was highest on nylon followed by wood and high-density polyethylene. The highest contact angle measurements were observed after 96-h of biofilm formation for each material. All the results and observations from this study contribute to a better understanding of the attachment and hydrophobicity characteristics of Salmonella and might help producers make informed decisions when selecting containers for harvesting and storing in order to minimize biofilm formation and potential for cross-contamination.

Stochastic processes shape microbial community assembly in biofilters: Hidden role of rare taxa

Stochastic and deterministic processes are the major themes governing microbial community assembly; however, their roles in bioreactors are poorly understood. Herein, the mechanisms underlying microbial assembly and the effect of rare taxa were studied in biofilters. Phylogenetic tree analysis revealed differences in microbial communities at various stages. Null model analysis showed that stochastic processes shaped the community assembly, and deterministic processes emerged only in the inoculated activated sludge after domestication. This finding indicates the dominant role of stochastic factors (biofilm formation, accumulation, and aging). The Sloan neutral model corroborated the advantages of stochastic processes and mainly attributed these advantages to rare taxa. Cooccurrence networks revealed the importance of rare taxa, which accounted for more than 85% of the keystones. Overall, these results provide good foundations for understanding community assembly, especially the role of rare taxa, and offer theoretical support for future community design and reactor regulation.

Faunal colonists, including mussel settlers, respond to microbial biofilms at deep-sea hydrothermal vents

Colonization processes at dynamic deep-sea hydrothermal vent ecosystems ultimately determine ecosystem structure, function, resilience, and recovery. Microbial biofilms form rapidly on surfaces near hydrothermal vents and are continuously exposed to the highly variable abiotic environment. Thus, biofilm microbes may provide a temporally integrated signal that can indicate whether the habitat is suitable for faunal colonists. This study explored the role of microbial biofilms in controlling faunal colonization through in-situ colonization experiments at Tica Vent in the 9°50’ N region of the East Pacific Rise (EPR). Short-term experiments (∼2 weeks) were conducted by deploying colonization surfaces (“sandwiches”) either with an established biofilm (developed for >1 year) or a fresh biofilm (developed throughout experiment) in zones characterized by different faunal assemblages. Differences in associated larval settlers, faunal immigrants, and microbial communities according to biofilm age across multiple biogenic zones were investigated. Faunal and microbial community compositions significantly differed according to whether the sandwiches had established or fresh biofilms as well as the biogenic zone they were deployed in. Several faunal colonists, including settlers such as the foundational chemosymbiotic mussel Bathymodiolus thermophilus and the nectochaete Archinome sp., were found associated more with established biofilms than fresh biofilms. Microbial biofilm communities were dominated by putative chemoautotrophic members of the Campylobacterota phylum and Gammaproteobacteria class and several microbial taxa were found to covary with faunal colonists. Overall, these findings show that microbial community composition plays a role in larval settlement and animal migration in hydrothermal vent systems and the detection of microbial and faunal interactions provides a starting point for identifying key microbial characteristics influencing colonization processes at hydrothermal vents.

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.

Lactic acid bacteria and spoilage bacteria: Their interactions in Escherichia coliO157:H7 biofilms on food contact surfaces and implications for beef contamination

AbstractThis research explores the interaction between Shiga toxin‐producing Escherichia coli (STEC) O157:H7 and bacteria species commonly found in beef processing environments, specifically Carnobacterium, Lactobacillus, Comamonas, Raoultella, and Pseudomonas. The study investigated how various environmental conditions impact the formation of biofilms and the ability of O157:H7 to transfer from multispecies biofilm onto beef surfaces. For this purpose, a mixture of lactic acid bacteria (LAB), spoilage bacteria (106 CFU/mL), and E. coli O157 (103 CFU/mL) were combined as follows: LAB (T1): Carnobacterium piscicola + Lactobacillus bulgaricus + O157:H7, an spoilage bacteria (T2): Comamonas koreensis + Raoultella terrigena + O157:H7, an spoilage bacteria (T3): Pseudomonas aeruginosa + C. koreensis strain + O157:H7 and only O157:H7 as control (T4). Multispecies biofilms were developed on thermoplastic polyurethane (TPU) and stainless steel (SS) coupons at 10 and 25°C for 6 days, washed and stored for 6, 30, and 60 days at wet (60%–90% RH) and dry (20%–50%, RH) conditions. To evaluate O157:H7 transfer, beef cubes (3 × 3 × 1 cm) were placed on the coupons, followed by a 50‐g weight (7.35 kPa). The experiment was repeated three times in triplicate for each strain combination. Results demonstrate that biofilms formed at 10°C were generally weaker (less biomass) than those at 25°C. Regardless of temperature, more viable O157:H7 cells were transferred to beef from moist biofilms on TPU surfaces. At 25°C, T3 biofilm exhibited the lowest O157:H7 transfer to beef by 1.44 log10 CFU/cm2 (p &lt; 0.01). At 10°C, none of the multispecies biofilm (T1–T3) affected the number of O157:H7 transfers to beef (p &gt; 0.05). Notably, O157:H7 was not detected on food contact surfaces with 30 and 60‐day‐old dry biofilms (T1–T4). Through enrichment, E. coli O157:H7 was recovered from multispecies biofilms T1, T2, and T3. Findings from this study imply that multispecies biofilms contribute to the persistence of O157:H7 under dry conditions, regardless of temperature. These results underscore the intricate influence of multiple environmental factors—including surface type, biofilm age, humidity, temperature, and the presence of other bacterial species—on the risk of beef contamination facilitated by biofilms.

Treatment of wound infections linked to neurosurgical implants.

As neurosurgery has advanced technologically, more and more neurosurgical implants are being employed on an aging patient population with several comorbidities. As a result, there is a steady increase in the frequency of infections linked to neurosurgical implants, which causes serious morbidity and mortality as well as abnormalities of the skull and inadequate brain protection. We discuss infections linked to internal and external ventricular and lumbar cerebrospinal fluid drainages, neurostimulators, craniotomies, and cranioplasty in this article. Biofilms, which are challenging to remove, are involved in all implant-associated illnesses. It takes a small quantity of microorganisms to create a biofilm on the implant surface. Skin flora bacteria are implicated in the majority of illnesses. Microorganisms that cause disruptions in wound healing make their way to the implant either during or right after surgery. In about two thirds of patients, implant-associated infections manifest early (within the first month after surgery), whereas the remaining infections present later as a result of low-grade infections or by direct extension from adjacent infections (per continuitatem) to the implants due to soft tissue damage. Except for ventriculo-atrial cerebrospinal fluid shunts, neurosurgical implants are rarely infected by the haematogenous route. This research examines established and clinically validated principles that are applicable to a range of surgical specialties using implants to treat biofilm-associated infections in orthopaedic and trauma cases. Nevertheless, there is little evidence and no evaluation in sizable patient populations to support the success of this extrapolation to neurosurgical patients. An optimal microbiological diagnostic, which includes sonicating removed implants and extending culture incubation times, is necessary for a positive result. Additionally, a strategy combining surgical and antibiotic therapy is needed. Surgical procedures involve a suitable debridement along with implant replacement or exchange, contingent on the biofilm's age and the state of the soft tissue. A protracted biofilm-active therapy is a component of antimicrobial treatment, usually lasting 4-12 weeks. This idea is appealing because it allows implants to be changed or kept in place for a single surgical procedure in a subset of patients. This not only enhances quality of life but also lowers morbidity because each additional neurosurgical procedure increases the risk of secondary complications like intracerebral bleeding or ischemia.

Improving green sea urchin (Strongylocentrotus droebachiensis) hatchery production by determining effective settlement cues and post-settlement conditions

Hatchery production methodologies of Northeastern U.S. green sea urchins (Strongylocentrotus droebachiensis) have yielded low settlement success and post-settlement survival (approximately 1–5%) contributing towards high costs for producing juveniles for this emerging aquaculture species. These are likely attributed to hatchery operations needing to optimize settlement cues and post-settlement food supplies. This study examined a range of potential settlement cues using adult conspecifics or a benthic diatom biofilm (Cylindrotheca closterium or Nitzschia sp.) alone, or in combination with seawater conditioned with conspecifics or macroalga (Saccharina latissima or Ulva lactuca). This study also examined whether sea urchin post-settlement survival and growth would improve using a benthic diatom food source (C. closterium or Nitzschia sp.) alone or in combination with an additional macroalga food source (S. latissima or U. lactuca). Both trials were conducted under hatchery ambient (control 12 °C) or elevated (14 °C) temperatures to determine whether temperature elevation could enhance S. droebachiensis settlement success and post-settlement growth. This study also examined what influence biofilm age (1, 4 and 8 weeks) and seawater salinity (26, 30, 33 and 36) had on settlement success to help towards optimizing hatchery methods. This study did not improve post-settlement survival causing it to remain a significant barrier in mass producing juveniles, but it did increase juvenile growth by 30–40% and reduce growth variability when fed the benthic diatom C. closterium in combination with U. lactuca. This combination also served as an adequate settlement cue for this species. The use of a conspecific influenced biofilm substantially improved settlement success from 1 to 5% to 50–80%. Seasonal timing and age (4 weeks) of this biofilm indicated importance in yielding this success which will be location specific. The use of conspecific biofilms for improving post-settlement survival requires further investigation and is worth examining in combination with C. closterium and U. lactuca towards enhancing S. droebachiensis production in the Northeastern U.S. Elevating seawater temperature by 2 °C had no significant effect on settlement, growth, or survival in this study. Salinities of 30–33 were optimal yielding 3–7 times more settlement compared to salinities 27 and 36. This study highlights that settlement trials need to incorporate post-settlement survival measurements for at least 1–2 months to truly reflect the effectiveness of method interventions and consider extending the time before turning seawater flow back on into settlement tanks from 24 to 48 h by up to 72–96 h to facilitate greater settlement success if seawater quality parameters permit it.

Assessing the impact of anti-microbial showerheads on the prevalence and abundance of opportunistic pathogens in shower water and shower water-associated aerosols

Respiratory infections from drinking water-associated pathogens that can cause infections in the immunocompromised (DWPIs) are increasing, yet knowledge of DWPI aerosolization and if dynamics are DWPI-specific is lacking. Although there are several DWPI mitigation strategies, the use of antimicrobial showerheads is one of the easiest and most economical. There are many manufacturers and designs of antimicrobial showerheads that claim to remove microorganisms from shower water, yet all fail to assess efficacy in realistic conditions. In this study, a custom-built shower laboratory housing triplicates of three different showerheads (antimicrobial filter-based, antimicrobial silver-embedded and conventional acrylonitrile butadiene styrene plastic) were used to assess the physiochemical and microbial dynamics in shower water and respirable shower water-associated aerosols (1µm – 5 µm) over the course of 84 days. Collectively, findings from the study suggest that showerheads marketed as antimicrobial produce similar chemical and DWPI water quality to non-antimicrobial showerheads (p= &amp;gt;0.05) when operated under real-world conditions, however marked differences in the rare microbial community were present. In addition, although there were no differences in absolute DWPI abundance between showerhead type, each DWPI peaked in concentration at a different biofilm ages, suggesting that potential DWPI inhalation risk is DWPI- specific and influenced by the number of days of operation of the showerhead.

Can you see the algae for the slime? Temporal patterns of biofilm food quality and quantity in lowland rivers

AbstractAnimals must invest some portion of their metabolism to activities related to physiological maintenance and the remainder to processes related to the production of new biomass for growth and reproduction. Animal metabolism is fuelled by food, and the quality and quantity of food, along with the effort invested to obtain it, are fundamental to supporting populations. Biofilms are a primary basal food resource within riverine food webs, and it is thought that their nutritional value for animals decreases with age due to dynamic changes in community composition. We sought to test assumptions of spatiotemporal changes to biofilm nutritional value by assessing variations in biofilm mass and fatty acid composition in three rivers for 73 days. We also used a multi‐prong eDNA approach to characterize changes to biofilm fungal (ITS1–4), bacterial (16S), and algal (23S) community compositions. We anticipated biofilm food value to decrease with biofilm age due to shifts in composition from high‐quality green algae and diatoms to low‐quality cyanobacteria and filamentous algae. Our results partially support this contention; biofilm food value, assessed as a combination of fatty acid mass per unit area (in grams per square meter) and concentration of fatty acids (in milligrams per gram), was dynamic and peaked between 24 and 43 days following submersion. After 43 days, biofilm food value decreased. However, despite significant temporal changes in biofilm community composition and a decrease in overall lipid concentration, the proportions of different fatty acid classes among total lipids did not vary. Instead, the observed increase in the abundance of cyanobacteria and filamentous algae compared with diatoms and green algae, along with higher quantities of lipid‐poor extracellular polymeric substances (EPS), likely contributed to the reduction in overall lipid concentration relative to the biofilm dry mass. Here we present a novel approach to balance consumer energetic costs with food quality within aquatic food webs. Our results have important implications for river management and provide valuable information for the use of environmental water to support lotic ecosystems.

Determinants of Microscale pH in In Situ-Grown Dental Biofilms.

Dental biofilm pH is the most important determinant of virulence for the development of caries lesions. Confocal microscopy-based pH ratiometry allows monitoring biofilm pH with high spatial resolution. Experiments performed on simplified biofilm models under static conditions identified steep pH gradients as well as localized acidogenic foci that promote enamel demineralization. The present work used pH ratiometry to perform a comprehensive analysis of the effect of whole saliva flow on the microscale pH in complex, in situ-grown 48-h and 96-h biofilms (n = 54) from 9 healthy participants. pH was monitored in 12 areas at the biofilm bottom and top, and saliva flow with film thicknesses corresponding to those in the oral cavity was provided by an additively manufactured microfluidic flow cell. Biofilm pH was correlated to the bacterial composition, as determined by 16S rRNA gene sequencing. Biofilm acidogenicity varied considerably between participants and individual biofilms but also between different areas inside one biofilm, with pH gradients of up to 2 units. pH drops were more pronounced in 96-h than in 48-h biofilms (P = 0.0121) and virtually unaffected by unstimulated saliva flow (0.8 mm/min). Stimulated flow (8 mm/min) raised average biofilm pH to near-neutral values but it did not equilibrate vertical and horizontal pH gradients in the biofilms. pH was significantly lower at the biofilm base than at the top (P < 0.0001) and lower downstream than upstream (P = 0.0046), due to an accumulation of acids along the flow path. pH drops were positively correlated with biofilm thickness and negatively with the thickness of the saliva film covering the biofilm. Bacterial community composition was significantly different between biofilms with strong and weak pH responses but not their species richness. The present experimental study demonstrates that stimulated saliva flow, saliva film thickness, biofilm age, biofilm thickness, and bacterial composition are important modulators of microscale pH in dental biofilms.

Assessing the health risks associated with the usage of water-atomization shower systems in buildings

In the context of climate change policies, buildings must implement solutions to reduce energy and water consumption. One such solution is showering with water atomization showerheads, which can significantly reduce water and energy usage. However, the lack of risk assessment for users' health has hindered the widespread adoption of this technology. To address this gap, we assess the risk of spreading bacteria, in particular the pathogenic bacterium Legionella pneumophila, from shower hose biofilms of different ages grown under controlled or uncontrolled conditions considering different levels of water hardness, during showering using water atomization showerheads (ECO) or continuous flow showerheads (STA). We compared the aerosol and bioaerosol emission – total, viable and cultivable – during a 10 min shower event between the two shower systems. We showed that the water-atomization showerhead emitted slightly more nanoparticles smaller than 0.45 µm and slightly fewer particles larger than 0.5 µm than the continuous flow showerhead. Additionally, ECO showerheads emitted fewer cultivable bacteria than STA, regardless of the biofilm's age or growth conditions. When Legionella pneumophila was detected in biofilms, ECO showerheads released slightly less cultivable Legionella in the first flush of shower water compared to the STA, ranging from 6.0 × 102 to 1.6 × 104 CFU·L−1. However, cultivable L. pneumophila was not detected in the aerosols emitted during showering with either showerhead. These findings suggest that emerging water-drop emission technologies might affect human exposure to aerosols differently than traditional systems, emphasizing the importance of assessing the health risks associated with any new shower system. Additionally, these findings provide valuable insights for achieving a balance between water and energy conservation.

Upregulation of ica Operon Governs Biofilm Formation by a Coagulase-Negative Staphylococcus caprae.

Staphylococcus caprae is a Gram-positive, coagulase-negative staphylococci (CoNS), which appears as commensal in the skin, as well as a prevalent mastitis pathogen of goats. Occasionally, it is also associated with infections in humans. Biofilm formation has been identified as a putative virulence factor in S. caprae. Biofilms are multicellular communities protected by a self-produced extracellular matrix (ECM), which facilitates the resistance of bacterial cells to antimicrobial treatments. The ECM is constructed by exopolysaccharides, including the major exopolysaccharide-polysaccharide intercellular adhesion (PIA), regulated by the ica operon in Staphylococcus species. The aim of this study was to characterize the expression of the ica operon in relation to biofilm formation in S. caprae. Results showed that within a few hours of growth, S. caprae could adhere to polystyrene surfaces, start to accumulate, and form biofilm. Peak biofilm biomass and maturation were reached after 48 h, followed by a reduction in biomass after 72 h. Confocal laser scanning microscopy showed the expression of matrix-associated proteins and polysaccharides at various time points. The expression dynamics of the ica operon were investigated using real-time reverse transcriptase PCR (RT)-qPCR, which showed elevated expression during the early stages of biofilm formation and subsequent downregulation throughout the biofilm aging process. In conclusion, our results show that the ica operon is essential in regulating biofilm formation in S. caprae, similar to other Staphylococcus species. Furthermore, the robustness of the observed biofilm phenotype could account for the successful intramammary colonization and may explain disease persistence caused by this pathogenic bacterium.

Comparative evaluation of the effectiveness of alcohol-based sanitizers, UV-C radiation and hot air on three-age Salmonella biofilms

Dry sanitation is recommended to control contamination and prevent microbial growth and biofilm formation in the low-moisture food manufacturing plants. The objective of this study was to evaluate the effectiveness of dry sanitation protocols on Salmonella three-age biofilms formed on stainless steel (SS) and polypropylene (PP). Biofilms were formed for 24, 48 and 96 h at 37 °C using a cocktail of six Salmonella strains (Muenster, Miami, Glostrup, Javiana, Oranienburg, Yoruba) isolated from the peanut supply chain. Then, the surfaces were exposed to UV-C radiation, hot air (90 °C), 70% ethanol and a commercial product based on isopropyl alcohol for 5, 10, 15 and 30 min. After 30min exposure, on PP the reductions ranged from 3.2 to 4.2 log CFU/cm2 for UV-C, from 2.6 to 3.0 log CFU/cm2 for hot air, from 1.6 to 3.2 log CFU/cm2 for 70% ethanol and from 1.5 to 1.9 log CFU/cm2 for the commercial product. On SS, after the same exposure time, reductions of 1.3–2.2 log CFU/cm2, 2.2 to 3.3 log CFU/cm2, 1.7 to 2.0 log CFU/cm2 and 1.6 to 2.4 log CFU/cm2 were observed for UV-C, hot air, 70% ethanol and commercial product, respectively. UV-C was the only treatment affected by the surface material (p < 0.05) whereas the biofilm age influenced the effectiveness of UV-C and hot air (p < 0.05). For most treatment, there was significant difference among the exposure times (p < 0.05). Overall, the fastest loss in the biofilm viability was noted in the first 5 min, followed by a tail phase. The time predicted by the Weibull model for the first decimal reduction ranged from 0.04 to 9.9 min on PP and from 0.7 to 8.5 min on SS. In addition, the Weibull model indicates that most of treatments (79%) required a long-term exposure time (>30 min) to achieve 3-log reductions of Salmonella biofilms. In summary, UV-C showed the best performance on PP whereas hot air was noted to be the most effective on SS.

The Heterogeneous Diffusion of Polystyrene Nanoparticles and the Effect on the Expression of Quorum-Sensing Genes and EPS Production as a Function of Particle Charge and Biofilm Age.

Biofilms are abundantly present in both natural and engineered environmental systems and will likely influence broader particle fate and transport phenomena. While some developed models describe the interactions between nanoparticles and biofilms, studies are only beginning to uncover the complexity of nanoparticle diffusion patterns. With the knowledge of the nanoparticle potential to influence bacterial processes, more systematic studies are needed to uncover the dynamics of bacteria-nanoparticle interactions. This study explored specific microbial responses to nanoparticles and the heterogeneity of nanoparticle diffusion. Pseudomonas aeruginosa biofilms (cultivated for 48 and 96 hours, representing early and late stages of development) were exposed to charged (aminated and carboxylated) polystyrene nanoparticles. With a combination of advanced fluorescence microscopy and real time quantitative PCR, we characterized the diffusion of polystyrene nanoparticles in P. aeruginosa biofilms and evaluated how biofilms respond to the presence of nanoparticles in terms of the expression of key EPS production-associated genes (pelA and rpsL) and quorum-sensing associated (lasR) genes. Our findings show that nanoparticle diffusion coefficients are independent of the particle surface charge only in mature biofilms and that the presence of nanoparticles influences bacterial gene expression. Independent of the particle's charge polystyrene nanoparticles down-regulated pelA in mature biofilms. By contrast, charge-specific responses were identified in lasR and rpsL gene expression. The targeted genes expression analysis and heterogeneous diffusion models demonstrate that particle charge influences nanoparticle mobility and provides significant insight into the intrinsic structural heterogeneity of P. aeruginosa biofilms. These findings suggest that biofilm maturity and particle charge are essential factors to consider when evaluating the transport of nanoparticles within a biofilm matrix.

Carbapenem-resistant biofilm-producing multidrug resistant Klebsiella pneumoniae induced hospital-acquired infections and analysis of resistant genes

In recent years, drug resistance has caused a significant number of fatalities. This review focuses on the emergence of medication resistance in Klebsiella pneumoniae strains associated with biofilm development, neonatal sepsis, aging, and secondary K. pneumoniae infection in COVID-19 cases. This review is an exclusive survey of &gt;200 research articles and &gt;100 review articles indexed in the Web of Science list with the keywords “K. pneumoniae,” “drug resistance,” “sepsis,” “neonatal sepsis,” “sepsis in older age,” “secondary infection,” “biofilm production,” “secondary infection in COVID-19,” “drug resistance genes,” and “whole genome sequencing”. In order to protect antibiotic susceptibility and to provide an overview of the biofilm forming mechanisms among gram-negative bacteria, the review discusses the function of multidrug-resistant K. pneumoniae in the production of biofilms. Additionally, K. pneumoniae’s role in contemporary cases of sepsis, spread of medication resistance in hospitals, and sepsis caused by K. pneumoniae in the geriatric age group are discussed. A summary of current approaches and the identification of drug resistance genes utilizing a whole genome sequencing technique are also reviewed in this article.

Electrochemical Characteristics of Shewanella loihica PV-4 on Reticulated Vitreous Carbon (RVC) with Different Potentials Applied.

The current output of an anodic bioelectrochemical system (BES) depends upon the extracellular electron transfer (EET) rate from electricigens to the electrodes. Thus, investigation of EET mechanisms between electricigens and solid electrodes is essential. Here, reticulated vitreous carbon (RVC) electrodes are used to increase the surface available for biofilm formation of the known electricigen Shewanella loihica PV-4, which is limited in conventional flat electrodes. S. loihica PV-4 utilizes flavin-mediated EET at potential lower than the outer membrane cytochromes (OMC), while at higher potential, both direct electron transfer (DET) and mediated electron transfer (MET) contribute to the current output. Results show that high electrode potential favors cell attachment on RVC, which enhances the current output. DET is the prevailing mechanism in early biofilm, while the contribution of MET to current output increased as the biofilm matured. Electrochemical analysis under starvation shows that the mediators could be confined in the biofilm. The morphology of biofilm shows bacteria distributed on the top layer of honeycomb structures, preferentially on the flat areas. This study provides insights into the EET pathways of S. loihica PV-4 on porous RVC electrodes at different biofilm ages and different set potential, which is important for the design of real-world BES.

Wetting and Imbibition Characteristics of Pseudomonas fluorescens Biofilms Grown on Stainless Steel.

This study aims to provide insights into biofilm resistance associated with their structural properties acquired during formation and development. On this account, the wetting and imbibition behavior of dehydrated Pseudomonas fluorescens biofilms grown on stainless steel electropolished substrates is thoroughly examined at different biofilm ages. A polar liquid (water) and a non-polar liquid (diiodomethane) are employed as wetting agents in the form of sessile droplets. A mathematical model is applied to appraise the wetting and imbibition performance of biofilms incorporating the evaporation of sessile droplets. The present results show that the examined biofilms are hydrophilic. The progressive growth of biofilms leads to a gradual increase of substrate surface coverage─up to full coverage─accompanied by a gradual decrease of biofilm surface roughness. It is noteworthy that just after 24 h of biofilm growth, the surface roughness increases about 6.7 times the roughness of the clean stainless steel surface. It is further found that the imbibition of liquid in the biofilm matrix is restricted only to the biofilm region under the sessile droplet. The lack of further capillary imbibition into the biofilm structure, beyond the droplet deposition region, implies that the biofilm matrix is not in the form of an extended network of interconnected micro/nanopores. All in all, the present results indicate a resilient biofilm structure to biocide penetration despite its hydrophilic nature.

Proteome of Staphylococcus aureus Biofilm Changes Significantly with Aging.

Staphylococcus aureus is a notorious biofilm-producing pathogen that is frequently isolated from implantable medical device infections. As biofilm ages, it becomes more tolerant to antimicrobial treatment leading to treatment failure and necessitating the costly removal of infected devices. In this study, we performed in-solution digestion followed by TMT-based high-throughput mass spectrometry and investigated what changes occur in the proteome of S. aureus biofilm grown for 3-days and 12-days in comparison with 24 h planktonic. It showed that proteins associated with biosynthetic processes, ABC transporter pathway, virulence proteins, and shikimate kinase pathway were significantly upregulated in a 3-day biofilm, while proteins associated with sugar transporter, degradation, and stress response were downregulated. Interestingly, in a 3-day biofilm, we observed numerous proteins involved in the central metabolism pathways which could lead to biofilm growth under diverse environments by providing an alternative metabolic route to utilize energy. In 12-day biofilms, proteins associated with peptidoglycan biosynthesis, sugar transporters, and stress responses were upregulated, whereas proteins associated with ABC transporters, DNA replication, and adhesion proteins were downregulated. Gene Ontology analysis revealed that more proteins are involved in metabolic processes in 3dwb compared with 12dwb. Furthermore, we observed significant variations in the formation of biofilms resulting from changes in the level of metabolic activity in the different growth modes of biofilms that could be a significant factor in S. aureus biofilm maturation and persistence. Collectively, potential marker proteins were identified and further characterized to understand their exact role in S. aureus biofilm development, which may shed light on possible new therapeutic regimes in the treatment of biofilm-related implant-associated infections.

Formation and Transfer of Multi-Species Biofilms Containing E. coli O103:H2 on Food Contact Surfaces to Beef.

Interactions of Shiga toxin–producing E. coli (STEC; O103:H2) with lactic acid bacteria (LAB) or spoilage bacteria (SP) multispecies biofilms on polyurethane (TPU) and stainless-steel (SS) were assessed at 10 and 25°C under wet and dry conditions after 6, 30, and 60 days of storage. One LAB T1: Carnobacterium piscicola + Lactobacillus bulgaricus, and two SP T2: Comamonas koreensis + Raoultella terrigena; T3: Pseudomonas aeruginosa + C. koreensis were assessed for their ability to form multispecies biofilms with O103:H2. O103:H2 single-species biofilms served as a control positive (T4). Coupons were stored dry (20–50% relative humidity; RH) or moist (60–90% RH) for up to 60 days, at which point O103:H2 transfer to beef and survival was evaluated. At 25°C, T3 decreased beef contamination with O103:H2 by 2.54 log10 CFU/g (P < 0.001). Overall, at 25°C contamination of beef with O103:H2 decreased (P < 0.001) from 3.17 log10 CFU/g on Day 6 to 0.62 log10 CFU/g on Day 60. With 60 days dry biofilms on TPU, an antagonistic interaction was observed among O103:H2 and multispecies biofilm T1 and T3. E. coli O103:H2 was not recovered from T1 and T3 after 60 days but it was recovered (33%) from T2 and T4 dry biofilms. At 10°C, contamination of beef with O103:H2 decreased (P < 0.001) from 1.38 log10 CFU/g after 6 days to 0.47 log10 CFU/g after 60 days. At 10°C, recovery of O103:H2 from 60 days dry biofilms could only be detected after enrichment and was always higher for T2 than T4 biofilms. Regardless of temperature, the transfer of O103:H2 to beef from the biofilm on TPU was greater (P < 0.001) than SS. Moist biofilms also resulted in greater (P < 0.001) cell transfer to beef than dry biofilms at 10 and 25°C. Development of SP or LAB multispecies biofilms with O103:H2 can either increase or diminish the likelihood of beef contamination. Environmental conditions such as humidity, contact surface type, as well as biofilm aging all can influence the risk of beef being contaminated by STEC within multi-species biofilms attached to food contact surfaces.

Biofilm growth characterization and treatment performance in a single stage partial nitritation/anammox process with a biofilm carrier

Biofilm carriers can avoid microorganism washout while maintaining a high amount of biomass, but are also associated with a long biofilm formation period and biofilm aging. A single stage partial nitritation/anammox process (single stage PN/A) reactor was setup to study the biofilm growth characterization and treatment performance under an NLR of 0.53 to 0.90 gN/L/d over one year. Biofilm growth was divided into three stages: the formation stage, maturation stage and aging stage. The initial biofilm was observed at day 84. A nitrogen removal efficiency of 83.4% was achieved at an NLR of 0.90 gN/L/d during the mature biofilm stage. Starvation, nitrogen gas accumulation and hydroxyapatite formation resulted in biofilm aging. After mechanical stirring treatment, biofilm reactivation was achieved by biofilm re-formation within one month. There is clear potential for phosphorus recovery, as indicated by the 5.24% - 6.29% phosphorus content in the biofilm (similar to the 5%-7% phosphorus content in enhanced biological phosphate removal sludge). The AnAOB genera abundance in the biofilm maintained at a high level of 18.25%-32.31%, while the abundance of AnAOB increased from the initial 4.10% to 13.78% after mechanical stirring treatment in the suspended sludge ensured biofilm reactivation. The results of this study clearly show that mechanical stirring treatment can be used to achieve the biofilm reactivation as the biofilm fills with the hollow cylindrical carrier. This study has potential as a useful reference for the realization of the wide application of the biofilm single stage PN/A process in the future.