Biofilm Detection Research Articles

Virulence characterization of Staphylococcus spp. isolated from mozzarella cheeses and cheese-slicers

Different species of the Staphylococcus genus can be found as contaminants in foods and may carry virulence factors that could potentially make them The enterotoxigenic potential ofpathogenic or transfer genes to other bacteria. Therefore, the aim of the present study was to isolate and identify species of Staphylococcus genus isolated from sliced mozzarella cheeses and cold-cut slicers and characterize biofilm formation in vitro, presence of enterotoxins and antibiotic resistance genes, and β-lactamase production. Sampling was carried out in 44 retail markets and isolates identified on species level. Resistance genes blaZ and mecA, as well as enterotoxin genes sea, seb, sec, and sed, were identified using PCR. Phenotypic evaluation of biofilm formation and detection of β-lactamase enzyme were also analyzed. Nine species of coagulase-negative Staphylococcus (CNS) were identified from 85 bacterial isolates. Of this total, 67.0% were positive for the presence of one or both antimicrobial resistance genes blaZ and mecA, while the β-lactamase enzyme was detected in 52.9% isolates. Regarding biofilm production, 48.2% of isolates were positive and 51.8% were negative. However, none of the isolates were identified with the enterotoxin genes. The results demonstrate the potential risk of cross-contamination, as biofilm-producing CNS could persist in the cheese retail environment, highlighting the danger of resistance genes transferring to other commensal or pathogenic bacteria.

Novel 3S-shaped biophotonic sensor utilizing MoS2–NSs/ZnO–NWs/AuCu–NCs for rapid detection of Shigella flexneri bacteria

This paper describes a unique, extremely sensitive biophotonic sensor with a three-tier S-tapered (3S) structure. It is designed for the real-time detection of Shigella flexneri (S. flexneri), a common foodborne pathogen that causes severe gastrointestinal diseases. The sensor development includes three distinct diameters of S-tapered structures. The performance of tapered sections was improved by using molybdenum disulfide nanosheets (MoS2-NSs), zinc oxide nanowires (ZnO-NWs), and photoluminescent bimetallic gold–copper nanoclusters (AuCu–NCs). These nanoparticles greatly improve the sensor’s performance. The sensor is further functionalized using anti-S. flexneri antibodies, allowing for the precise detection and capture of the target bacterium. The results show that the sensor can detect S. flexneri rapidly and accurately, with a linear detection range of 1–108 colony-forming units per milliliter (CFU/ml) and a low detection limit of 4.412 CFU/ml. In addition, the sensor’s ability to identify S. flexneri biofilms is demonstrated. Biofilm detection allows us to better understand and control biofilm concerns in the environment, equipment, and biomedical devices. Aptamer examines confirm the sensor’s ability to detect S. flexneri from the lateral direction. This study makes a significant contribution to the field of biosensing because no biophotonic sensor has previously been developed specifically for the detection of S. flexneri, fulfilling a critical gap in the arena of food safety and pathogen detection. The 3S sensor’s performance, robustness, and potential for practical applications make it an important addition to the field of photonics.

A Review of Bacterial Biofilm Components and Formation, Detection Methods, and Their Prevention and Control on Food Contact Surfaces

The microbial biofilms are a community of microorganisms that adhere to each other and to surfaces, typically in a mucilaginous or gel-like matrix composed of extracellular polymeric substances, including polysaccharides, proteins, lipids, and DNA. In the food industry, the bacterial biofilms may be formed on different surfaces and cause post-processing contamination or cross-contamination from the food contact surfaces to food products. Conventional cleaning and sanitizing methods are often ineffective at removing bacterial biofilms. Among more recent alternative methods proposed to address this problem are the use of hydrolytic enzymes, essential oils, and bacteriocins. These methods show promise since their antibacterial and antibiofilm actions involve degradation of the extracellular polymeric matrix of the biofilm and lead to inhibition of the foodborne pathogens present. Understanding the limitations and mechanisms of action of enzymes, bacteriocins, and essential oils in controlling bacterial biofilms on foods and food contact surfaces is essential for developing solutions to prevent and control biofilm formation. This review critically summarizes the current knowledge of bacterial biofilm components, their formation, detection methods, prevention, and removal from food contact surfaces.

Optimizing Diagnosis and Management of Ventilator-Associated Pneumonia: A Systematic Evaluation of Biofilm Detection Methods and Bacterial Colonization on Endotracheal Tubes

Healthcare-associated infections, such as ventilator-associated pneumonia and biofilm formation on intubation cannulas, impose significant burdens on hospitals, affecting staffing, finances, and patient wellbeing, while also increasing the risk of patient mortality. We propose a research study aimed at exploring various methodologies for detecting these infections, discovered in the biofilm on medical devices, particularly tracheal cannulas, and understanding the role of each method in comprehending these infections from an etiological perspective. Our investigation also involves an analysis of the types of endotracheal tubes utilized in each case, the bacteria species identified, and strategies for combating biofilm-associated infections. The potential impact of our research is the substantial improvement of patient care through enhanced diagnosis and management of these infections.

Assessing Biofilm at the Bedside: Exploring Reliable Accessible Biofilm Detection Methods.

Biofilm is linked through a variety of mechanisms to the pathogenesis of chronic wounds. However, accurate biofilm detection is challenging, demanding highly specialized and technically complex methods rendering it unapplicable for most clinical settings. This study evaluated promising methods of bedside biofilm localization, fluorescence imaging of wound bacterial loads, and biofilm blotting by comparing their performance against validation scanning electron microscopy (SEM). In this clinical trial, 40 chronic hard-to-heal wounds underwent the following assessments: (1) clinical signs of biofilm (CSB), (2) biofilm blotting, (3) fluorescence imaging for localizing bacterial loads, wound scraping taken for (4) SEM to confirm matrix encased bacteria (biofilm), and (5) PCR (Polymerase Chain Reaction) and NGS (Next Generation Sequencing) to determine absolute bacterial load and species present. We used a combination of SEM and PCR microbiology to calculate the diagnostic accuracy measures of the CSB, biofilm blotting assay, and fluorescence imaging. Study data demonstrate that 62.5% of wounds were identified as biofilm-positive based on SEM and microbiological assessment. By employing this method to determine the gold truth, and thus calculate accuracy measures for all methods, fluorescence imaging demonstrated superior sensitivity (84%) and accuracy (63%) compared to CSB (sensitivity 44% and accuracy 43%) and biofilm blotting (sensitivity 24% and accuracy 40%). Biofilm blotting exhibited the highest specificity (64%), albeit with lower sensitivity and accuracy. Using SEM alone as the validation method slightly altered the results, but all trends held constant. This trial provides the first comparative assessment of bedside methods for wound biofilm detection. We report the diagnostic accuracy measures of these more feasibly implementable methods versus laboratory-based SEM. Fluorescence imaging showed the greatest number of true positives (highest sensitivity), which is clinically relevant and provides assurance that no pathogenic bacteria will be missed. It effectively alerted regions of biofilm at the point-of-care with greater accuracy than standard clinical assessment (CSB) or biofilm blotting paper, providing actionable information that will likely translate into enhanced therapeutic approaches and better patient outcomes.

Application of Online Flow Cytometry for Early Biofouling Detection in Reverse Osmosis Membrane Systems.

Biofouling poses a significant challenge to reverse osmosis (RO) membrane systems, necessitating timely detection for effective control. This study evaluated the efficacy of flow cytometry (FCM) for early biofilm detection in comparison to conventional system performance indicators. Feed channel pressure drop and total cell concentration in the Membrane Fouling Simulator (MFS) flowcell cross-flow outlet water were monitored over time as early biofouling indicators. The results demonstrated the potential of increased bacterial cell concentration in cross-flow outlet water as a reliable indicator of biofouling development on the membrane. Water outlet monitoring enabled faster biofouling detection compared to feed channel pressure drop. Membrane autopsy confirmed biofilm presence prior to the pressure drop increase, highlighting the advantage of early detection in implementing corrective measures. Timely intervention reduces operational costs and energy consumption in membrane-based processes.

Correlation between Klebsiella Species’ Biofilm Formation and Antibiotic Resistance from Erbil Hospital Patients

The primary cause of nosocomial infections is Klebsiella pneumonia, an environmental gram-negative, encapsulated, non-motile bacterium. Antibiotic resistance and biofilm formation are two crucial characteristics that contribute to the pathogenicity of this bacterium. This research aimed to examine the relationship between antibiotic resistance and the capability of Klebsiella species isolated from Erbil hospital patients to produce biofilms. Therefore, in this study, 38 samples were collected from different laboratories and hospitals. At first, the samples were rejuvenated in Tryptic soy broth and then cultured on MacConkey agar. Gram staining and urease test were performed to identify the bacterial isolates while VITEK 2 system was performed to determine both the bacterial isolates and the antibiotic susceptibility. At last, Biofilm detection was assessed by the microliter plate technique as well as the Congo red agar method. The results revealed positive Klebsiella samples that were both resistant to some antibiotics and biofilm producers.

Artificial Intelligence-Driven Analysis of Antimicrobial-Resistant and Biofilm-Forming Pathogens on Biotic and Abiotic Surfaces.

The growing threat of antimicrobial-resistant (AMR) pathogens to human health worldwide emphasizes the need for more effective infection control strategies. Bacterial and fungal biofilms pose a major challenge in treating AMR pathogen infections. Biofilms are formed by pathogenic microbes encased in extracellular polymeric substances to confer protection from antimicrobials and the host immune system. Biofilms also promote the growth of antibiotic-resistant mutants and latent persister cells and thus complicate therapeutic approaches. Biofilms are ubiquitous and cause serious health risks due to their ability to colonize various surfaces, including human tissues, medical devices, and food-processing equipment. Detection and characterization of biofilms are crucial for prompt intervention and infection control. To this end, traditional approaches are often effective, yet they fail to identify the microbial species inside biofilms. Recent advances in artificial intelligence (AI) have provided new avenues to improve biofilm identification. Machine-learning algorithms and image-processing techniques have shown promise for the accurate and efficient detection of biofilm-forming microorganisms on biotic and abiotic surfaces. These advancements have the potential to transform biofilm research and clinical practice by allowing faster diagnosis and more tailored therapy. This comprehensive review focuses on the application of AI techniques for the identification of biofilm-forming pathogens in various industries, including healthcare, food safety, and agriculture. The review discusses the existing approaches, challenges, and potential applications of AI in biofilm research, with a particular focus on the role of AI in improving diagnostic capacities and guiding preventative actions. The synthesis of the current knowledge and future directions, as described in this review, will guide future research and development efforts in combating biofilm-associated infections.

Challenges in Biofilm Identification in Diabetic Foot Infections: Review of Literature.

Foot ulcerations are one of the most common complications of diabetes and one of the major initial causes of amputations. The formation of biofilms on wounds significantly contributes to infections and delayed healing. While existing methods for identifying these biofilms have limitations, there is a need for a convenient tool for its clinical application. This literature review aimed to address the problem with current clinical biofilm identification in wound care and a proposal for biofilm-detection-based wound care in diabetic foot ulcer patients. Identifying biofilms is particularly vital due to the absence of typical signs of infection in DFUs. However, current approaches, although effective, often prove invasive and technically intricate. The wound blotting technique, involving attaching a nitrocellulose membrane and subsequent staining, presents an alternative that is swift and non-invasive. Research highlights the applicability of wound blotting with alcian blue staining in clinical scenarios, consistently producing sensitive outcomes. By addressing the critical need for early biofilm detection, wound blotting holds promise for enhancing DFU management and contributing to strategies aimed at preventing amputations.

Biofilm Production and Its Implications in Pediatrics.

Biofilms, aggregates of bacteria enclosed in a self-produced matrix, have been implicated in various pediatric respiratory infections, including acute otitis media (AOM), otitis media with effusion (OME), adenoiditis, protracted bacterial bronchitis, and pulmonary exacerbations in cystic fibrosis. These infections are prevalent in children and often associated with biofilm-producing pathogens, leading to recurrent and chronic conditions. Biofilms reduce antibiotic efficacy, contributing to treatment failure and disease persistence. This narrative review discusses biofilm production by respiratory pathogens such as Streptococcus pneumoniae, non-typeable Haemophilus influenzae, Pseudomonas aeruginosa, and Staphylococcus aureus. It examines their mechanisms of biofilm formation, antibiotic resistance, and the challenges they present in clinical treatment. Various antibiofilm strategies have shown promise in vitro and in animal studies, including the use of N-acetylcysteine, enzymes like dispersin B, and agents disrupting quorum sensing and biofilm matrix components. However, their clinical application, particularly in children, remains limited. Traditional treatments for biofilm-associated diseases have not significantly evolved, even with biofilm detection. The transition from experimental findings to clinical practice is complex and requires robust clinical trials and standardized biofilm detection protocols. Addressing biofilms in pediatric respiratory infections is crucial for improving treatment outcomes and managing recurrent and chronic diseases effectively.

Determine Biofilm Genes in Pseudomonas Aeruginosa Isolated from Clinical and Environmental Samples

Objective: Isolates of Pseudomonas aeruginosa are an extremely adaptable bacterium that causes opportunistic diseases because of its varied metabolic pathways, genes, virulence factors, and considerable antibiotic resistance. Methods: A total of 293 samples were collected from different places: 193 samples (% 65.87) of human samples and 100 samples (% 34.13) of wastewater samples in the period between 3rd September to 15th November 2023). Bacterial isolates were identified according to microscopic, cultural, and genetic characteristics. Antibiotic susceptibility of bacterial isolates was determined against twelve of the selected antibiotics. The biofilm production was done by using phenotypic ways (Congo red agar and Microtiter plate methods) as well as genotypic ways by detection of biofilm genes (algD, pelf, and pslD genes). Results: Hundred-forty eight bacterial isolates were obtained, and sixty of these isolates were identified as Pseudomonas spp. (40.9%), twenty-six isolates of E. coli (17.9%), seventeen isolates of K. pneumoniae (11.3%), and forty-five isolates were beyond to other types of bacteria (30.1%), and out of sixty isolates of Pseudomonas spp., forty-two were identified as P. aeruginosa isolates. P. aeruginosa isolates revealed various resistance levels to antimicrobial agents gradually, ranging from 83.87% to Trimethoprim-sulfamethoxazole (SXT) to 3.22% to Aztreonam (AZT). Biofilm production by using the Congo red method showed that 27 isolates (64.28%) were positive results, while in the microtiter method, all forty-two isolates were positive (100%), the genetic detection showed that the AlgD gene was recognized in thirty-one isolates (73.8%), followed by Pelf and PslD genes in four isolates each (9.5%). Conclusion: The isolation percentage showed a high occurrence of multi-drug resistance biofilm forming Pseudomonas spp. isolates which could be a critical indicator. Methods of biofilm detection showed that the microtiter plate method has accuracy more than the Congo red method; as well AlgD gene was prevalent compared with both other genes Pelf, and PslD.

Detection of MRSA and MSSA biofilms in clinical specimens from a tertiary care hospital located in Hyderabad

Detection of MRSA and MSSA biofilms in clinical specimens from a tertiary care hospital located in Hyderabad

Real-time biofilm detection techniques: advances and applications.

Microbial biofilms, complex assemblies enveloped in extracellular matrices, are significant contributors to various infections. Traditional in vitro biofilm characterization methods, though informative, often disrupt the biofilm structure. The need to address biofilm-related infections urgently emphasizes the importance of continuous monitoring and timely interventions. This review provides a focused examination of advancements in real-time biofilm detection techniques, specifically in electrochemical, optical and mechanical systems. The potential applications of real-time detection in managing and monitoring biofilm growth in industrial settings, preventing medical infections, comprehending biofilm dynamics and evaluating control strategies highlight the necessity for it. Crucially, the review emphasizes the importance of evaluating these methods for their accuracy and reliability in real-time biofilm detection, offering valuable insights for precise interventions across various applications.

Development of super nanoantimicrobials combining AgCl, tetracycline and benzalkonium chloride

In this work, we demonstrate that a simple argentometric titration is a scalable, fast, green and robust approach for producing AgCl/antibiotic hybrid antimicrobial materials. We titrated AgNO3 into tetracycline hydrochloride (TCH) aqueous solution, thus forming AgCl/TCH in a one-step procedure. Furthermore, we investigated the one-pot synthesis of triply synergistic super-nanoantimicrobials, combining an inorganic source of Ag+ ions (AgCl), a disinfecting agent (benzyl-dimethyl-hexadecyl-ammonium chloride, BAC) and a molecular antibiotic (tetracycline hydrochloride, TCH). Conventional antimicrobial tests, industrial biofilm detection protocols, and in situ IR-ATR microbial biofilm monitoring, have been adapted to understand the performance of the synthesized super-nanoantimicrobial. The resulting hybrid AgCl/BAC/TCH nanoantimicrobials are found to be synergistically active in eradicating Salmonella enterica and Lentilactobacillus parabuchneri bacteria and biofilms. This study paves the way for the development of a new class of super-efficient nanoantimicrobials that combine relatively low amounts of multiple active species into a single (nano)formulation, thus preventing the development of antimicrobial resistance towards a single active principle.

Biofilm-Producing and Specific Antibiotic Resistance Genes in Pseudomonas aeruginosa Isolated from Patients Admitted to a Tertiary Care Hospital, Bangladesh

Pseudomonas aeruginosa is one of the organisms well-known for producing biofilm. Biofilms are responsible for persistent infections and antimicrobial resistance. The aim of this was to investigate P. aeruginosa for its ability to form biofilm. Genes that were responsible for the production of biofilms and biofilm-specific antimicrobial resistance were detected. The association between antibiotic resistance and biofilm was investigated. This cross-sectional study was conducted from July 2017 to December 2018. A total of 446 samples (infected burns, surgical wounds, and ETA) were collected from admitted patients at Dhaka Medical College and Hospital, Bangladesh. P. aeruginosa was isolated and identified by biochemical tests and PCR. Biofilm production by the tissue culture plate (TCP) method was followed by the detection of biofilm[1]producing genes (pqsA, pslA, pslD, pslH, pelA, lasR) and biofilm-specific antibiotic resistance genes (ndvB, PA1874, PA1876, PA1877) by PCR. The antibiotic susceptibility test was carried out by the disc diffusion method; for colistin agar dilution, the MIC method was followed. Among 232 (52.02%) positive strains of P. aeruginosa, 24 (10.30%) produced biofilms in TCPM. Among biofilm-producing genes, pqsA was found the highest number of isolates (79.17%), which was followed by pslA and pelA (70.83%). Other were found in lesser extent. Among the biofilm-specific antibiotic resistance genes, 16.67% of the isolates had ndvB, and 8.33% had PA1874 and PA1877. Biofilm-forming strains were significantly resistant to colistin in comparison to non-biofilm-forming ones. In conclusion, detection of biofilm-forming genes may be a good tool for the evaluation of biofilm production, which will help in prompt and better management of chronic or device-associated infections. Bangladesh J Microbiol, Volume 40, Number 2, December 2023, pp 60-65

A Comprehensive Review of Microbial Biofilms on Contact Lenses: Challenges and Solutions.

Contact lenses (CL) have become an immensely popular means of vision correction, offering comfort to millions worldwide. However, the persistent issue of biofilm formation on lenses raises significant problems, leading to various ocular complications and discomfort. The aim of this review is to develop safer and more effective strategies for preventing and managing microbial biofilms on CL, improving the eye health and comfort of wearers. Taking these into consideration, the present study investigates the intricate mechanisms of biofilm formation, by exploring the interplay between microbial adhesion, the production of extracellular polymeric substances, and the properties of the lens material itself. Moreover, it emphasizes the diverse range of microorganisms involved, encompassing bacteria, fungi, and other opportunistic pathogens, elucidating their implications within lenses and other medical device-related infections and inflammatory responses. Going beyond the challenges posed by biofilms on CL, this work explores the advancements in biofilm detection techniques and their clinical relevance. It discusses diagnostic tools like confocal microscopy, genetic assays, and emerging technologies, assessing their capacity to identify and quantify biofilm-related infections. Finally, the paper delves into contemporary strategies and innovative approaches for managing and preventing biofilms development on CL. In Conclusion, this review provides insights for eye care practitioners, lens manufacturers, and microbiology researchers. It highlights the intricate interactions between biofilms and CL, serving as a foundation for the development of effective preventive measures and innovative solutions to enhance CL safety, comfort, and overall ocular health. Research into microbial biofilms on CL is continuously evolving, with several future directions being explored to address challenges and improve eye health outcomes as far as CL wearers are concerned.

Simulation of a Radio-Frequency Wave Based Bacterial Biofilm Detection Method in Dairy Processing Facilities

This paper describes the principles behind the radio-frequency (RF) sensing of bacterial biofilms in pipes and heat exchangers in a dairy processing plant using an electromagnetic simulation. Biofilm formation in dairy processing plants is a common issue where the absence of timely detection and subsequent cleaning can cause serious illness. Biofilms are known for causing health issues and cleaning requires a large volume of water and harsh chemicals. In this work, milk transportation pipes are considered circular waveguides, and pasteurizers/heat exchangers are considered resonant cavities. Simulations were carried out using the CST studio suite high-frequency solver to determine the effectiveness of the real-time RF sensing. The respective dielectric constants and loss tangents were applied to milk and biofilm. In our simulation, it was observed that a 1 µm thick layer of biofilm in a milk-filled pipe shifted the reflection coefficient of a 10.16 cm diameter stainless steel circular waveguide from 0.229 GHz to 0.19 GHz. Further sensitivity analysis revealed a shift in frequency from 0.8 GHz to 1.2 GHz for a film thickness of 5 µm to 10 µm with the highest wave reflection (S11) peak of ≈−120 dB for a 6 µm thick biofilm. A dielectric patch antenna to launch the waves into the waveguide through a dielectric window was also designed and simulated. Simulation using the antenna demonstrated a similar S11 response, where a shift in reflection coefficient from 0.229 GHz to 0.19 GHz was observed for a 1 µm thick biofilm. For the case of the resonant cavity, the same antenna approach was used to excite the modes in a 0.751 m × 0.321 m × 170 m rectangular cavity with heat exchange fins and filled with milk and biofilm. The simulated resonance frequency shifted from 1.52 GHz to 1.54 GHz, for a film thickness varying from 1 µm to 10 µm. This result demonstrated the sensitivity of the microwave detection method. Overall, these results suggest that microwave sensing has promise in the rapid, non-invasive, and real-time detection of biofilm formation in dairy processing plants.

Microrobotics in endodontics: A perspective.

Microorganisms are the primary aetiological factor of apical periodontitis. The goal of endodontic treatment is to prevent and eliminate the infection by removing the microorganisms. However, microbial biofilms and the complex root canal anatomy impair the disinfection process. Effective and precise endodontic therapy could potentially be achieved using advanced multifunctional technologies that have the ability to access hard-to-reach surfaces and perform simultaneous biofilm killing, removal, and detection of microorganisms. Advances in microrobotics are providing novel therapeutic and diagnostic opportunities with high precision and efficacy to address current biofilm-related challenges in biomedicine. Concurrently, multifunctional magnetic microrobots have been developed to overcome the disinfection challenges of current approaches to disrupt, kill, and retrieve biofilms with the goal of enhancing the efficacy and precision of endodontic therapy. This article reviews the recent advances of microrobotics in healthcare and particularly advances to overcome disinfection challenges in endodontics, and provides perspectives for future research in the field.

Biofilm monitoring through the detection of cyclic diguanosine-monophosphate with an easy-to-use electrochemical sensor

The identification and monitoring of biofilm is of utmost importance because it ensures the survival of bacteria in a variety of environments, such as nature, medical indwelling devices, hospital surfaces, or the food industry. Here, we describe the novel application of a screen-printed electrode modified with nanomaterials for the in situ detection of bacterial biofilm. This approach enables simple, rapid, and highly sensitive and selective electrochemical detection of cyclic diguanosine-monophosphate (cdGMP). The best electrochemical signal of cdGMP was obtained after testing several electrode materials and screen-printed electrodes modified with different nanomaterials, different electrolyte solutions, pH, and electrochemical technique parameters. Using the anodic peak of the cdGMP on screen-printed electrodes modified with carbon nanotubes (CNT-SPE), the sensor showed excellent sensitivity, with a 25 nM–1 µM linear range, and a limit of detection of 15 nM. The sensor allowed the selective detection of cdGMP, with low interference from other molecules and was successfully applied to real sample analyses (laboratory biofilm produced by various bacterial species over distinct time intervals).

In vitro ANALYSIS OF BIOFILM FORMATION AND ANTIBIOTIC RESISTANCE OF UROPATHOGENIC Escherichia coli

Urinary tract infections are an exceedingly common worldwide problem, caused mostly by Gram-negative bacteria, especially Escherichia coli. Microbial biofilms are considered a serious public health problem. The potential of uropathogenic E. coli (UPEC) to produce biofilm was explored in the present study. The Congo red agar, tube- and tissue culture plate methods were used to evaluate the formation of biofilm by E. coli isolates. Of the 155 isolates, 101, 106 and 90 isolates were positive for these three methods, respectively. Subsequently, the sensitivity of E. coli isolates to antimicrobial agents (amikacin, cefepime, cefixime, cefotaxime, cefpodoxime, ceftazidime, ceftriaxone, ciprofloxacin, nitrofurantoin, gentamycin, nalidixic acid, ofloxacin and pefloxacin) was tested. There was no difference in the rate of biofilm detection between Congo red agar method and tube method. The antibiotic sensitivity test revealed that the biofilm producing isolates were multi-drug resistant. The study emphasized the necessity for developing alternative therapeutic approaches to overcome multi drug resistance arising from biofilm formation of UPEC.