Biofilm System Research Articles

Enrichment of phosphate-accumulating organisms (PAOs) in a microfluidic model biofilm system by mimicking a typical aerobic granular sludge feast/famine regime

Wastewater treatment using aerobic granular sludge has gained increasing interest due to its advantages compared to conventional activated sludge. The technology allows simultaneous removal of organic carbon, nitrogen, and phosphorus in a single reactor system and is independent of space-intensive settling tanks. However, due to the microscale, an analysis of processes and microbial population along the radius of granules is challenging. Here, we introduce a model system for aerobic granular sludge on a small scale by using a machine-assisted microfluidic cultivation platform. With an implemented logic module that controls solenoid valves, we realized alternating oxic hunger and anoxic feeding phases for the biofilms growing within. Sampling during ongoing anoxic cultivation directly from the cultivation channel was achieved with a robotic sampling device. Analysis of the biofilms was conducted using optical coherence tomography, fluorescence in situ hybridization, and amplicon sequencing. Using this setup, it was possible to significantly enrich the percentage of polyphosphate-accumulating organisms (PAO) belonging to the family Rhodocyclaceae in the community compared to the starting inoculum. With the aid of this miniature model system, it is now possible to investigate the influence of a multitude of process parameters in a highly parallel way to understand and efficiently optimize aerobic granular sludge-based wastewater treatment systems.Key points• Development of a microfluidic model to study EBPR.• Feast-famine regime enriches polyphosphate-accumulating organisms (PAOs).• Microfluidics replace sequencing batch reactors for aerobic granular sludge research.

Persistent Inhibition of Ammonium Released from Contaminated Sediments Through a Modified Zeolite and Biofilm System Enhanced by Signaling Molecules

This experiment used a modified zeolite and biofilm system to find a long-term effective way of repairing sediment. Four types of modified zeolites[AlCl3, Al(NO3)3, Al2(SO4)3, and KAl(SO4)2] and a biofilm system were investigated for the removal of ammonia nitrogen from overlaying water. The results showed that, of the modified zeolites, AlCl3 had the greatest effect on the zeolite and biofilm system, and the optimal modified concentration was 0.8 mol·L-1. The persistence was further studied after adding OHHL, including the inhibition of ammonium released from contaminated sediment by the AlCl3-modified zeolite and biofilm system, NaCl-modified zeolite and biofilm system, and natural zeolite and biofilm system. The results illuminated that the number of bacteria attached to the AlCl3-modified zeolite and biofilm system was the highest, and the proportion of denitrifying bacteria was also high (the initial proportion was 82.1%; 30 days later it was 61.1%). Therefore, the biological regeneration was high (64.9%), which caused the zeolite service life to be up to 8.5 months. Thus, ammonium released from contaminated sediment can be inhibited effectively and continuously by an AlCl3-modified zeolite and biofilm system.

Technical Note 8 - Applicability of support medium based of polyurethane foam for systems used to post-treatment of anaerobic effluents

Several technologies have been applied to post-treat anaerobic effluents. For most of those, a rigorous sludge management is needed. In this case, the use of sponge-based packing media in biofilm systems tends to significantly contribute to the minimization of sludge production and effluent quality improvement. Endogenous respiration and predation are important mechanistic processes explaining such improvements, which is related with the increase of the sludge retention time by the retention of biomass within the sponge. Depending on the operational condition, nitrogen removal is also observed in addition to the notorious performance for organic matter removal. Considering the advances for the post-treatment of anaerobic effluents with the use of sponge-based biofilm systems, this Technical Note aim to present: (i) fundamentals of biofilm systems using polyurethane sponge as support media; (ii) full-scale experiences with emerging sponge-based technologies; (iii) performance of sponge-based biofilm systems and operational conditions usually practiced; (iv) a preliminary critical analysis regarding implementation and operational costs; (v) advantages and disadvantages. The potential and limitations regarding the use of sponge-based reactors as post-treatment of anaerobic effluents are finally remarked.

Bacterial Cellulose Membrane as Carrier Support for High-Biomass Production of Polyhydroxyalkanoates from Food Waste in a Biofilm System

Bacterial Cellulose Membrane as Carrier Support for High-Biomass Production of Polyhydroxyalkanoates from Food Waste in a Biofilm System

Fabrication of Wood-Plastic Composites as Biocarriers in Moving Bed Biofilm Reactors

In present study, a novel biocarrier was fabricated from wood and high density polyethylene (HDPE) using a single screw extruder. The effect of wood:plastic mass ratio on physico-chemical properties of the obtained wood plastic composite (WPC) was evaluated. While the tensile and flexural strengths were decreased, the sample density was increased with higher wood contents. The wood:plastic mass ratio of 1:1 was considered as the best for the production of WPC biocarriers. The role of this material in moving bed biofilm system was tested in a batch mode reactor having volume of 2 L with 20% filling by the biocarrier. The treatment efficiencies of organic matter, ammonia and phosphate were enhanced nearly two times. The results are attributed to heterogenous and porous surface of WPC, which enhances bacteria adhesion and pollutant removal. It demonstrates that WPC can be a potential biocarrier for use in moving bed biofilm reactors (MBBRs).

Phosphorus recovery in the alternating aerobic/anaerobic biofilm system: Performance and mechanism

To balance the high phosphorus concentration in recirculated solution and the stability of biofilm system, this study explored the performance and mechanism of phosphorus uptake/release for recovering phosphorus from sewage when the phosphorus content in biofilm (Pbiofilm) changed. The results showed that the maximum phosphorus concentration in the concentrated solution reached 171.2 ± 2.5 mg·L−1 in harvest 1st-5th stages. Polyphosphate accumulating organisms (PAOs) performed a metabolic shift from glycogen accumulation metabolism (GAM) to polyphosphate accumulation metabolism (PAM) when Pbiofilm increased at each phosphorus enrichment stage, and more phosphorus was absorbed/released by PAOs. Nevertheless, the release of poly-phosphate from PAOs was inhibited after phosphorus concentration stabilized, and PAOs were unable to absorb phosphorus from wastewater as it reached the phosphorus saturation stage. To maintain the stability of the system, phosphorus had to be harvested so that the saturated phosphorus in PAOs was easily released in a new recirculated solution, resulting in adequate storage space for PAOs to absorb phosphorus. Meanwhile, the 31P NMR analysis demonstrated that phosphorus was stored in EPS and cell of PAOs, whereas EPS played a significant role than cell at the anaerobic phase. Particularly, ortho-phosphate was the major component of phosphorus release by EPS and poly-phosphate was the major part of phosphorus release by cell. Furthermore, the change of Pbiofilm had no impact on biofilm characteristics and microbial communities, whereas some PAOs would be enriched, and others that were not suitable for this process would be inhibited with repeated cycles of alternating aerobic/anaerobic operation.

Biofilm Biology and Engineering of Geobacter and Shewanella spp. for Energy Applications.

Geobacter and Shewanella spp. were discovered in late 1980s as dissimilatory metal-reducing microorganisms that can transfer electrons from cytoplasmic respiratory oxidation reactions to external metal-containing minerals. In addition to mineral-based electron acceptors, Geobacter and Shewanella spp. also can transfer electrons to electrodes. The microorganisms that have abilities to transfer electrons to electrodes are known as exoelectrogens. Because of their remarkable abilities of electron transfer, Geobacter and Shewanella spp. have been the two most well studied groups of exoelectrogens. They are widely used in bioelectrochemical systems (BESs) for various biotechnological applications, such as bioelectricity generation via microbial fuel cells. These applications mostly associate with Geobacter and Shewanella biofilms grown on the surfaces of electrodes. Geobacter and Shewanella biofilms are electrically conductive, which is conferred by matrix-associated electroactive components such as c-type cytochromes and electrically conductive nanowires. The thickness and electroactivity of Geobacter and Shewanella biofilms have a significant impact on electron transfer efficiency in BESs. In this review, we first briefly discuss the roles of planktonic and biofilm-forming Geobacter and Shewanella cells in BESs, and then review biofilm biology with the focus on biofilm development, biofilm matrix, heterogeneity in biofilm and signaling regulatory systems mediating formation of Geobacter and Shewanella biofilms. Finally, we discuss strategies of Geobacter and Shewanella biofilm engineering for improving electron transfer efficiency to obtain enhanced BES performance.

Community-level and function response of photoautotrophic periphyton exposed to oxytetracycline hydrochloride

Periphyton is considered important for removal of organic pollutants from water bodies, but knowledge of the impacts of antibiotics on the community structure and ecological function of waterbodies remains limited. In this study, the effects of oxytetracycline hydrochloride (OTC) on the communities of photoautotrophic epilithon and epipelon and its effect on nitrogen and phosphorus concentrations in the water column were studied in a 12-day mesocosm experiment. The dynamics of nitrogen and phosphorus concentrations in the epipelon and epilithon experiment showed similar patterns. The concentrations of total nitrogen, dissolved total nitrogen, ammonium nitrogen, total phosphorus and dissolved total phosphorus in the water column increased rapidly during the initial days of exposure, after which a downward trend occurred. In the epilithon experiment, we found that the photosynthesis (Fv/Fm) and biomass of epilithon were significantly (P < 0.05) stimulated in the low concentration group. Contrarily, growth and photosynthesis (Fv/Fm) were significantly (P < 0.05) reduced in the medium and high concentration group. We further found that the photosynthetic efficiency of photoautotrophic epilithon was negatively correlated with the concentrations of nitrogen and phosphorus in the water column (P < 0.05). Principal coordinate analysis (PCoA) showed that the communities of epilithic algae in the control group and in the low concentration group were significantly (P < 0.05) different from that of the high concentration group during the initial 4 days. After 8 days’ exposure, all groups tended to be similar, indicating that epilithon showed rapid adaptability and/or resilience. Similar results were found for the relative abundance of some epilithic algae. Our findings indicate that the biofilm system has strong tolerance and adaptability to OTC as it recovered fast after an initial suppression, thus showing the important role of periphyton in maintaining the dynamic balance of nutrients with other processes in aquatic ecosystems.

Nitrogen and phosphorus removal efficiency and algae viability in an immobilized algae and bacteria symbiosis system with pink luminescent filler

Abstract In this study, an immobilized algae and bacteria symbiotic biofilm reactor (ABSBR) with pink luminescent filler (PLF) was constructed. The effects of PLF addition in the construction of an algae and bacteria symbiotic biofilm system on the nitrogen and phosphorus removal efficiencies and algae viability were evaluated. Our results showed that for influent TN and TP concentrations of 40 ± 5 and 5 ± 0.8 mg/L, respectively, the pollutant removal rates (PRRs) of TN and TP by the ABSBR can reach up to 74.74% and 88.36%, respectively. The chlorophyll-a (chl-a) concentration on the PLF reaches approximately 5,500 μg/L with a specific oxygen generation rate (SOGR) of 65.48 μmolO2 mg−1Chl-a h−1. These results indicate that the adding PLF into algae and bacteria symbiosis systems can effectively improve the nitrogen and phosphorus removal efficiencies of the sewage as well as increase biomass and viability of the algae in the system.

Up-regulation of ribosomal and carbon metabolism proteins enhanced pyrene biodegradation in fulvic acid-induced biofilm system

The polycyclic aromatic hydrocarbons (PAHs) that enter the aqueous phase usually coexist with fulvic acid (FA). Therefore, we initiated this investigation to explore the influences of FA on bacterial biofilm formation and its potential to biodegrade pyrene (PYR), using electron microscopic techniques and isobaric tags for relative and absolute quantification (iTRAQ). Our results revealed that FA stimulated biofilm formation and enhanced the biodegradation of PYR. First, FA favored the three-dimensional proliferation of bacteria, with an OD590/OD600 value of up to 14.78, and the extracellular surfaces covered by a layer of biomaterials. Distinctive intracellular morphologies of texture and organization were accompanied by reduced inter-bacterial distances of less than 0.31 μm. The biofilms formed displayed interactions between FA and surficial proteins, as noted by band shifts for the C–O and CO groups. Strikingly, FA triggered the upregulation of 130 proteins that were either operational in biofilm formation or in metabolic adjustments; with the changes supported by the increasing intensity of free amino acids and the newly generated N–O bonds. The results above revealed that the enhanced biodegradation was related to the up-regulation of the proteins functioned for ribosomal and carbon metabolism, and the ultra-structural changes in FA-induced biofilm system.

Role of carrier characteristics affecting microbial density and population in enhanced nitrogen and phosphorus removal from wastewater

This research aims to improve simultaneous nitrification-denitrification and phosphorus removal (SNDPR) using novel carriers and to demonstrate the effect of carrier characteristics on nutrient removal in a biofilm reactor. For this purpose, biofilms enriched with both polyphosphate-accumulating organisms (PAOs) and nitrifiers were cultivated in two parallel sequencing batch reactors containing conventional moving bed bioreactor carriers (MBBR) and a novel type of carriers (carbon-based moving carriers (CBMC)). The new carriers were produced based on recycled waste materials via a chemical-thermal process and their specific surface area were 10.4 times higher than typical MBBR carriers of similar dimensions. The results showed that the use of CBMC carriers increased bacterial adhesion by about 18.5% and also affected the microbial population inside the biofilms, leading to an increase in PAOs abundancy and thus an increase in biological phosphorus removal up to 12.5%. Additionally, it was corroborated that the volume of the anoxic zones with dynamic behavior is strictly influenced by the carrier structure and biofilm thickness due to a limitation in oxygen penetration. Accordingly, the formation of broader anoxic zones and shrinkage of these zones to a lesser extent resulted in the continuation of anoxic reactions for longer periods using the novel carriers. Thereby, an increase in nitrogen removal by about 15% was obtained mainly by denitrifying PAOs. The results also exhibited that a higher simultaneous nitrification-denitrification (SND) efficiency can be achieved by selecting an appropriate aeration program influencing the dynamic changes of anoxic zones. Overall, a biofilm system using the new carriers, with phosphorus and nitrogen removal efficiencies of 97.5% and 92.3%, was presented as an efficient, compact, and simple operation SNDPR process.

In-situ fermentation coupling with partial-denitrification/anammox process for enhanced nitrogen removal in an integrated three-stage anoxic/oxic (A/O) biofilm reactor treating low COD/N real wastewater

Mainstream partial-denitrification with anammox (PD-anammox) process faced the challenge of complex organics involved in real sewage. Herein, PD-anammox coupled with in-situ fermentation was successfully achieved in a full biofilm system formed by three-stage anoxic/oxic reactor to treat real wastewater with low COD/N of 3.6. The total nitrogen (TN) removal efficiency was enhanced to 78.4% ± 3.6% with average TN and ammonium concentrations in effluent of 10.6 and 0.5 mg N/L, respectively. Batch tests confirmed that partial-denitrification was the major nitrite provider for anammox in the anoxic biofilm, while in-situ fermentation could decompose the complex organics to readily-biodegradable organics for full- or partial-denitrification. Additionally, a significant anammox bacteria (Candidatus Brocadia) population was detected in the second (3.53%) and third (4.46%) anoxic zones, while denitrifiers and fermentative bacteria were mainly enriched in the first anoxic zone. This study presents a feasible approach for PD-anammox process in practical application under mainstream condition.

Long-term effect of fulvic acid amendment on the anammox biofilm system at 15 ℃: performance, microbial community and metagenomics analysis

The role of fulvic acid (FA) on the anammox system at 15 ℃ was investigated. After operation for 113 days, total inorganic nitrogen removal efficiency in FA amendment reactor achieved to 58.6% on average, higher than that of control group (42.1%). Anammox-related functional genes, i.e., hzo and hzs, also demonstrated higher expression level after introduction of FA. It was observed that Candidatus Kuenenia became more competitive than Candidatus Brocadia with the existence of FA at 15 ℃. Also, co-occurrence analysis showed that FA stimulated the complexity and interactive relationship of microbial communities in the anammox system. Metagenomics analysis revealed that FA introduction stimulated relative abundances of genes in central pathway of tricarboxylic acid cycle such as ACO, IDH, OGDH, SCS, FUM, and MDH. Meanwhile, metabolomics analysis revealed that metabolites related to amino sugar metabolic pathways (glucose 1-phosphate, UDP-D-glucuronate, UDP) and redox reactions (NAD+ and NADH) improved in the FA amendment reactor.

High-temperature biofilm system based on heterotrophic nitrification and aerobic denitrification treating high-strength ammonia wastewater: Nitrogen removal performances and temperature-regulated metabolic pathways

Conventional autotrophic nitrification process is difficult to treat high-temperature wastewater with high-strength ammonia. In this study, a high-temperature (50 °C) biofilm system based on heterotrophic nitrification and aerobic denitrification (HNAD) was established. The results showed that the HNAD process was high temperature resistant, and the nitrogen removal performance, pathway and microbial mechanism varied remarkably at different temperatures. The high-temperature system showed excellent nitrogen and COD removal capacities at 50 °C. Ammonia oxidation was mainly undertaken by heterotrophic nitrification, while anoxic and aerobic pathways worked in concert for denitrification. High-throughput sequencing indicated that heterotrophic nitrifying bacteria (8.58%) and denitrifying bacteria (52.88%) were dominant at 50 °C. Metagenomic analysis further suggested that the carbon metabolism was up-regulated in response to the increasing temperature, so more energy was generated, thereby promoting the HNAD-related nitrogen removal pathways. The study revealed the microbial mechanism of HNAD at high temperature and provided new insights into high-temperature biological nitrogen removal.

The combination of ultrasound and chlorogenic acid to inactivate Staphylococcus aureus under planktonic, biofilm, and food systems

This study aimed to investigate the mechanism of different treatments, namely, ultrasound (US), chlorogenic acid (CA), and ultrasound combined with chlorogenic acid (US plus CA) on the inactivation of Staphylococcus aureus planktonic and biofilm cells. Results showed that the combined treatment of US and CA exhibited remarkable synergistic antibacterial and antibiofilm effects. Scanning electron microscopy images indicated that the combined treatment of US and CA caused the most serious damage to the cell morphology. Confocal laser scanning microscopy images revealed that the combined treatment led to sharp increase and severe damage to the permeability of the cell membrane, causing the release of ATP and nucleic acids and decreasing the exopolysaccharide contents in S. aureus biofilm. Finally, the combined treatment of US plus 1% CA for 60 min inactivated S. aureus cells by 1.13 lg CFU/g on mutton. Thus, the combined treatment of US and CA had synergistic effect against S. aureus under planktonic, biofilm, and food systems.

A Review of Microalgal Biofilm Technologies: Definition, Applications, Settings and Analysis

Biofilm-based algal cultivation has many advantages over the conventional suspended growth methods and has received increased attention as a potential platform for algal production, wastewater treatment (nutrient removal), and a potential pathway to supply feedstock for microalgae-based biorefinery attempts. However, the attached cultivation by definition and application is a result of a complex interaction between the biotic and abiotic components involved. Therefore, the entire understanding of the biofilm nature is still a research challenge due to the need for real-time analysis of the system. In this review, the state of the art of biofilm definition, its life cycle, the proposed designs of bioreactors, screening of carrier materials, and non-destructive techniques for the study of biofilm formation and performance are summarized. Perspectives for future research needs are also discussed to provide a primary reference for the further development of microalgal biofilm systems.

Evaluation of the efficiency of an integrated biofilm and phytoremediation system with Nasturtium officinale for the treatment of municipal wastewater in Huancavelica

In this research removal efficiency was evaluated of organic pollutants of a biofilm and phytoremediation integrated system for municipal wastewater treatment in Huancavelica. A pilot plant consisting of a settler was installed that also retained oils and fats, a biological filter using as support material the calcareous stuff and a phytoremediation process with Nasturtium officinale (watercress) The hydraulic retention time was 5.5 hours and the flowrate of 0.011 L s -1, the evaluation was made of the proper functioning of processes by monitoring control parameters. The pollutant removal efficiency was determined by Student's "t" test (p &lt;0.05). An average pH value of 7.57 and a temperature of 12.07 ° C were obtained, subsequently the measurement of BOD5 was initiated, obtaining maximum effluent values of 131 ppm and minimum values of 45 ppm and a removal efficiency of 44.8% was obtained, for COD. In total coliforms, fecal coliforms and Escherichia coli the removal was 66.67%, 94.59%, 81.22% and 72.12% respectively, oil and fat analysis was also performed and total suspended solids, however high removal efficiency was not obtained. It is concluded that the parameters of BOD5 and COD comply with the MPL for the effluents of municipal domestic wastewater treatment plants, while the rest of the parameters analyzed do not comply with the regulations. however high removal efficiency was not obtained. It is concluded that the parameters of BOD5 and COD comply with the MPL for the effluents of municipal domestic wastewater treatment plants, while the rest of the parameters analyzed do not comply with the regulations. however high removal efficiency was not obtained. It is concluded that the parameters of BOD5 and COD comply with the MPL for the effluents of municipal domestic wastewater treatment plants, while the rest of the parameters analyzed do not comply with the regulations.

B-Cell Deficiency Exacerbates Inflammation and Bone Loss in Ligature-Induced Experimental Periodontitis in Mice.

ObjectivePeriodontitis, one of the most prevalent chronic oral infectious diseases in humans, is induced by the breakdown in the balance between the biofilm and host immune system. Previous studies have shown the presence of large numbers of B cells in periodontitis lesions, implicating that B lymphocytes play a predominant role during the pathogenesis of periodontitis. This study aimed to investigate the role of all B cells in the initiation of periodontitis.MethodsExperimental periodontitis was induced in B cell-deficient (CD19Cre) mice and wild-type (WT) control mice by 5-0 silk ligation around the maxillary second molar. Four weeks after ligation, alveolar bone loss was determined by micro-computed tomography. The levels of inflammatory cytokines and receptor activator of NF-κB ligand (RANKL)/osteoprotegerin in periodontal lesions were analyzed using real-time quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemistry. Lymphocyte populations in the cervical lymph nodes and spleen and among the peripheral blood mononuclear cells were detected by flow cytometry.ResultsB-cell deficiency resulted in increased severity of alveolar bone loss in mouse experimental periodontitis, which was associated with increased osteoclast activity and upregulated RANKL expression in the periodontal lesions. In addition, gingiva cytokine expression profiles were shifted to T helper type 1 (Th1) and Th17 in the CD19Cre mice with ligature-induced periodontitis compared with WT mice. In addition, a reduced CD4+/CD8+ T cell ratio was observed in the CD19Cre mice.ConclusionB-cell deficiency exacerbates the inflammation and alveolar bone loss in ligature-induced experimental periodontitis in mice, implicating that B cells may overall play a protective role in the initiation of periodontitis.

Isolation of Total RNA from Pseudomonas aeruginosa within Biofilms for Measuring Gene Expression.

Pseudomonas aeruginosa is an opportunistic bacterial pathogen that causes infections in the airways of cystic fibrosis (CF) patients. P. aeruginosa is known for its ability to form biofilms that are protected by a matrix of exopolysaccharides. This matrix allows the microorganisms to be more resilient to external factors, including antibiotic treatment. One of the most common methods of biofilm growth for research is in microtiter plates or chambered slides. The advantage of these systems is that they allow for the testing of multiple growth conditions, but their disadvantage is that they produce limited amounts of biofilm for RNA extraction. The purpose of this article is to provide a detailed, step by step protocol on how to extract total RNA from small amounts of biofilm of sufficient quality and quantity for high throughput sequencing. This protocol allows for the study of gene expression within these biofilm systems.

No Correlation between Biofilm Formation, Virulence Factors, and Antibiotic Resistance in Pseudomonas aeruginosa: Results from a Laboratory-Based In Vitro Study.

Pseudomonas aeruginosa (P. aeruginosa) possesses a plethora of virulence determinants, including the production of biofilm, pigments, exotoxins, proteases, flagella, and secretion systems. The aim of our present study was to establish the relationship between biofilm-forming capacity, the expression of some important virulence factors, and the multidrug-resistant (MDR) phenotype in P. aeruginosa. A total of three hundred and two (n = 302) isolates were included in this study. Antimicrobial susceptibility testing and phenotypic detection of resistance determinants were carried out; based on these results, isolates were grouped into distinct resistotypes and multiple antibiotic resistance (MAR) indices were calculated. The capacity of isolates to produce biofilm was assessed using a crystal violet microtiter-plate based method. Motility (swimming, swarming, and twitching) and pigment-production (pyoverdine and pyocyanin) were also measured. Pearson correlation coefficients (r) were calculated to determine for antimicrobial resistance, biofilm-formation, and expression of other virulence factors. Resistance rates were the highest for ceftazidime (56.95%; n = 172), levofloxacin (54.97%; n = 166), and ciprofloxacin (54.64%; n = 159), while lowest for colistin (1.66%; n = 5); 44.04% (n = 133) of isolates were classified as MDR. 19.87% (n = 60), 20.86% (n = 63) and 59.27% (n = 179) were classified as weak, moderate, and strong biofilm producers, respectively. With the exception of pyocyanin production (0.371 ± 0.193 vs. non-MDR: 0.319 ± 0.191; p = 0.018), MDR and non-MDR isolates did not show significant differences in expression of virulence factors. Additionally, no relevant correlations were seen between the rate of biofilm formation, pigment production, or motility. Data on interplay between the presence and mechanisms of drug resistance with those of biofilm formation and virulence is crucial to address chronic bacterial infections and to provide strategies for their management.