Detection of Biofilm Formation by Different Bacterial Isolates of Contact Lens

2020 ◽  
Vol EJMM29 (4) ◽  
pp. 93-100
Author(s):  
Wageih S. El. Naghy ◽  
Sarah A. Hamam ◽  
Tamer A. Wasfy ◽  
Sara M. Samy
Keyword(s):  
Staphylococcus Aureus ◽  
Contact Lens ◽  
Congo Red ◽  
Biofilm Formation ◽  
Detection Methods ◽  
Coagulase Negative Staphylococcus ◽  
Bacterial Isolates ◽  
Biofilm Production ◽  
Agar Method ◽  
Icaa Gene

Background: Biofilms are groups of microorganisms that collect to each other and with different surfaces by adherence mechanisms. These are formed of cells and extracellular matrix manufactured by these cells. There may be a great problem in some situations e.g. on medical implants and resistance against antibiotics. Objective: The objective of this study is to determine biofilm forming power of bacteria isolated from the conjunctiva, contact lens and the lens storage case by both phenotypic and genotypic detection methods. Methodology: Samples were taken from (36) persons in the period from January 2020 to June 2020 at Ophthalmology Department, Tanta University Hospitals, all the samples were transported to the Medical Microbiology & Immunology Department, Tanta University where bacterial strains were isolated. The biofilm formation phenotypic detection was performed by both tube method and Congo red agar method. The biofilm-forming genes of coagulase negative Staphylococcus (CoNS) and Staphylococcus aureus (ica A) and that of P. aeruginosa (psl A), were detected by PCR. Results: The (216) samples (swabs & discarded lenses) gave rise to a total number of (247) bacterial isolates. By using tube method; (52.3%) were moderately positive, (31.5%) strongly positive and (16.2%) negative for biofilm formation while after using the Congo red agar method; (35.3%) were moderately positive, (38.4%) strongly positive and (26.3%) negative for biofilm formation. Regarding the Staphylococcus aureus isolates, two (50%) of these were containing (icaA) gene. Regarding the (21) CoNS isolates, three (14.3%) contained (icaA) gene. Although all of the Pseudomonas isolates didn't contain pslA (1119 bp) gene, these were positive for biofilm production by phenotypic methods. Conclusion: The majority of the isolates had the capacity to form biofilms. Both tube and Congo red agar methods showed clear significant correlation and detected a high number of biofilm-producing strains. The absence of genes responsible for biofilm formation did not exclude the phenotypic biofilm production by these bacteria which is a common state.

scholarly journals Comparison of Microbiological Examination by Test Tube and Congo Red Agar Methods to Detect Biofilm Production on Clinical Isolates

2018 ◽  
Vol 54 (1) ◽  
pp. 22
Author(s):  
Dewi Klarita Furtuna ◽  
Kartuti Debora ◽  
Eddy Bagus Warsito
Keyword(s):  
Medical Devices ◽  
Congo Red ◽  
Biofilm Formation ◽  
Test Tube ◽  
Patient Treatment ◽  
Assay Method ◽  
Simple Method ◽  
Microtiter Plate Assay ◽  
Biofilm Production ◽  
Agar Method

Biofilm on medical devices can cause significant diseases and deaths and give a large effecton disease transmission among patients and health providers and potentially increasethe cost of patient treatment. By knowing the presence of biofilm on a patient, one can differentiate the treatment management for that particular patient from the patients without biofilm on their medical device. The purpose of this study was to obtain diagnostic method to detect biofilm formation on isolates from the medical devices by simple method that is easy to do and can be applied in resource-limited microbiology laboratory. 36 specimens obtained from IV Line, CVC, urinary catheter and ETT were grown on Muller Hinton agar and continued with 3 methods, i.e., Test Tube method, Congo Red Agar method and Microtiter Plate Assay method. Results of this study showed Test Tube (nephelometer), Test Tube (visual) and Congo Red Agar in order to have the same sensitivity of 100% but has higher specificity compared to Test Tube method (visual) and Congo Red Agar method in detecting biofilm production on isolates from medical devices that had been plugged into patients body. The biofilm formation inside devices depends on factors, i.e., host, device and the microorganism itself.

scholarly journals Biofilm Formation by Staphylococcus aureus Isolated from Food Contact Surfaces in the Dairy Industry of Jalisco, Mexico

2018 ◽  
Vol 2018 ◽  
pp. 1-8 ◽  
Author(s):  
María-Guadalupe Avila-Novoa ◽  
Maricarmen Iñíguez-Moreno ◽  
Oscar-Alberto Solís-Velázquez ◽  
Jean-Pierre González-Gómez ◽  
Pedro-Javier Guerrero-Medina ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Stainless Steel ◽  
Congo Red ◽  
Biofilm Formation ◽  
Dairy Industry ◽  
Food Contact ◽  
Biofilm Production ◽  
Food Contact Surfaces ◽  
Food Borne ◽  
Contact Surfaces

Staphylococcus aureus is an important food-borne pathogen able to form biofilms. This pathogen is responsible for outbreaks of food-borne illnesses associated with the consumption of milk and dairy products. The aim of this study was to evaluate the biofilm-forming ability of S. aureus isolates, recovered from food contact surfaces in the dairy industry of Jalisco, Mexico. A total of 84 S. aureus strains were evaluated. The isolates were characterized phenotypically by culture on Congo red agar plates. The ability of the strains to form biofilms was investigated in 96-well flat-bottomed microtiter polystyrene plates. Stainless-steel coupons were used as an experimental surface. Biofilm formation was observed, using epifluorescence microscopy and scanning electron microscopy. Detection of the icaADBC genes in S. aureus was performed by the PCR technique. A total of 52.3% (44/84) of the S. aureus strains contained the icaADBC gene that synthesizes polysaccharide intercellular adhesion (PIA) molecules. On Congo red agar, 75% (63/84) of the S. aureus isolates were biofilm producers, 16.6% (14/84) were non-biofilm formers, and 8.3% (7/84) showed a noncharacteristic phenotype. The biofilm production of the S. aureus strains SA-4E, SA-9, SA-13, and SA-19 on stainless-steel coupons was investigated at 25°C for 8 days, and the detected cell population density was approximately 7.15–7.82 log CFU cm−2. In addition to the ability of biofilm production, it is important to highlight that these strains are potential enterotoxin producers as se genes have been previously detected in their genomes. A part of the ability of biofilm production and the determination of the presence of virulence determinants in the genome of S. aureus can contribute to the pathogenicity of strains. Therefore, vigilant food safety practices need to be implemented in the dairy industries regarding FCS to prevent food-borne infections and intoxications due to S. aureus contamination.

scholarly journals Paradigm Shift in Antimicrobial Resistance Pattern of Bacterial Isolates during the COVID-19 Pandemic

Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 954
Author(s):  
Vikas Saini ◽  
Charu Jain ◽  
Narendra Pal Singh ◽  
Ahmad Alsulimani ◽  
Chhavi Gupta ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Tertiary Care ◽  
Public Health Problem ◽  
Resistance Pattern ◽  
Coagulase Negative Staphylococcus ◽  
Care Hospital ◽  
Bacterial Isolates ◽  
Hospital Infection Control ◽  
And Staphylococcus Aureus

Antimicrobial resistance (AMR) is an emerging public health problem in modern times and the current COVID-19 pandemic has further exaggerated this problem. Due to bacterial co-infection in COVID-19 cases, an irrational consumption of antibiotics has occurred during the pandemic. This study aimed to observe the COVID-19 patients hospitalized from 1 March 2019 to 31 December 2020 and to evaluate the AMR pattern of bacterial agents isolated. This was a single-center study comprising 494 bacterial isolates (blood and urine) that were obtained from patients with SARS-CoV-2 admitted to the ICU and investigated in the Department of Microbiology of a tertiary care hospital in Delhi, India. Out of the total bacterial isolates, 55.46% were gram negative and 44.53% were gram positive pathogens. Of the blood samples processed, the most common isolates were CoNS (Coagulase Negative Staphylococcus) and Staphylococcus aureus. Amongst the urinary isolates, most common pathogens were Escherichia coli and Staphylococcus aureus. A total of 60% MRSA was observed in urine and blood isolates. Up to 40% increase in AMR was observed amongst these isolates obtained during COVID-19 period compared to pre-COVID-19 times. The overuse of antibiotics gave abundant opportunity for the bacterial pathogens to gradually develop mechanisms and to acquire resistance. Since the dynamics of SARS-COV-2 are unpredictable, a compromise on hospital antibiotic policy may ultimately escalate the burden of drug resistant pathogens in hospitals. A shortage of trained staff during COVID-19 pandemic renders it impossible to maintain these records in places where the entire hospital staff is struggling to save lives. This study highlights the extensive rise in the use of antibiotics for respiratory illness due to COVID-19 compared to antibiotic use prior to COVID-19 in ICUs. The regular prescription audit followed by a constant surveillance of hospital infection control practices by the dedicated teams and training of clinicians can improve the quality of medications in the long run and help to fight the menace of AMR.

scholarly journals Molecular Investigation of Fibronectin-Binding Protein Genes and Capacity of Biofilm Production in Staphylococcus Aureus Isolated From Clinical Samples

2021 ◽  
Author(s):  
Hossein Jafari Soghondicolaei ◽  
Mohammad Ahanjan ◽  
Mehrdad Gholami ◽  
Bahman Mirzaei ◽  
Hamid Reza Goli
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Colorimetric Assay ◽  
Binding Protein ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Biofilm Production ◽  
Fibronectin Binding Protein ◽  
Fibronectin Binding

Abstract Biofilm production increases Staphylococcus aureus resistance to antibiotics and also host defense mechanisms. The current study aims to evaluate the biofilm formation by S. aureus and to determine the prevalence of fibronectin-binding protein genes, also its correlation with drug resistance. In this study, 100 clinical isolates of S. aureus were collected. The antibiotic susceptibility pattern of the isolates was evaluated by the disk agar diffusion method. The ability of biofilm formation in the studied isolates was also determined by microplate colorimetric assay. Then, all isolates were screened by polymerase chain reaction for the fnbA and fnbB genes. Out of 100 clinical isolates of S. aureus, the highest and lowest antibiotic resistance rates were against penicillin (94%) and vancomycin (6%). Thirty-two cases were found to be multi-drug resistant (MDR) among the all strains. The ability of biofilm production was observed in 89% of the isolates. The PCR results showed that the prevalence of fnbA and fnbB genes were 91% and 17%, respectively. Moreover, 100% and 21.8% of the MDR strains harbored the fnbA and fnbB genes respectively. The ability to form biofilm in MDR isolates of S. aureus is more than non-MDR isolates, especially fnbA positive ones. As the bacteria in the biofilm are difficult to kill by antibiotics, attention to the removal or control of the biofilm production seems to be necessary.

scholarly journals Evaluation of the Biofilm Forming Capacities of Bacterial Isolates Recovered in Raw and Treated Effluent from Wastewater Treatment Plant of Ahmadu Bello University Zaria, Nigeria

2019 ◽  
Vol 23 (10) ◽  
pp. 1783-1786
Author(s):  
MI Ugwoke ◽  
DA Machido ◽  
MB Tijjani
Keyword(s):  
Wastewater Treatment ◽  
Congo Red ◽  
Antimicrobial Agents ◽  
Wastewater Treatment Plants ◽  
Screening Method ◽  
Treatment Plant ◽  
Bacterial Isolates ◽  
Biofilm Production ◽  
Treated Effluent ◽  
Raw Wastewater

Biofilm producing bacteria are associated with many recalcitrant infections and are highly resistant to antimicrobial agents, hence notoriously difficult to eradicate. This study aimed at determining the biofilm forming capacities of bacterial isolates recovered in the raw wastewater and treated effluent from Wastewater Treatment Plants of Ahmadu Bello University Zaria using Tube Method (TM) and Congo Red Agar (CRA) method; and from the results, among the isolates recovered from the raw wastewater, TM detected 62.5% isolates as positive and 37.5% as negative for biofilm production, CRA detected 37.5% isolates as positive and 62.5% as negative for biofilm production. TM also demonstrated to be more suitable in detecting biofilm producing bacterial isolates from the treated effluent were it detected 50% isolates as positive and 50% as negative. However, CRA detected only 12.5% isolates as positive and 87.5% as negative for biofilm production. We therefore, conclude that the TM is more efficient and reliable for detection of biofilm producing bacteria in the laboratory when compared to CRA method and can be recommended as one of the suitable standard screening method for the detection of biofilm producing bacteria in laboratories.Keywords: Biofilm; Bacteria; Congo red agar and Tube method

Identification of ica-dependent biofilm production by Staphylococcus aureus clinical isolates and antibiofilm effects of ascorbic acid against biofilm production

2020 ◽  
Vol 73 (5) ◽  
pp. 261-266
Author(s):  
Sahra Kırmusaoğlu ◽  
Havva Kaşıkçı
Keyword(s):  
Staphylococcus Aureus ◽  
Ascorbic Acid ◽  
Biofilm Formation ◽  
Surface Proteins ◽  
Clinical Isolates ◽  
High Morbidity ◽  
Dependent Manner ◽  
Independent Manner ◽  
Biofilm Production ◽  
Life Threatening

AimsStaphylococcus aureus (S. aureus) is a life-threatening pathogen with high morbidity and mortality rates which causes nosocomial and community-acquired infections. Biofilm, considered to be a common virulence factor for pathogens, plays a significant role in recurrent and untreatable infections. Biofilm formation of S. aureus is mediated by synthesis of either poly-N-acetylglucosamine in an ica-dependent manner or surface proteins in an ica-independent manner. In some cases treatment is impossible and recurrent. In this study, ica-dependent biofilm-producing S. aureus isolates were detected and the anti-biofilm effect of ascorbic acid against biofilm formation of isolates was investigated.MethodsA total of 21 methicillin-sensitive S. aureus (MSSA) clinical isolates stored in our bacterial stock were used to detect ica-dependent biofilm-producing MSSA isolates. The anti-biofilm study was undertaken with three ica-dependent biofilm-producing isolates (MSSA2–4) and ATCC 29213 (MSSA1). Biofilms and the anti-biofilm effect of ascorbic acid were detected using the microtitre plate (MtP) method. 16S-rRNA, nuc, icaA and icaD genes and expression levels of icaA and icaD of isolates were detected by RT-PCR.ResultsThe minimum inhibitory concentrations (MICs) of ascorbic acid prevented biofilm formation of MSSA1 and MSSA3. Also, 1/2 MIC of ascorbic acid prevented biofilm formation of MSSA3. It was observed that biofilm formation decreased with increased concentration. There was no significant increase in ica gene expression of MSSA1 and MSSA2. Expression of icaA and icaD of MSSA3 decreased 13% and 38%, respectively. Expression of icaA in MSSA4 decreased 12%.ConclusionThe results of our study show that ascorbic acid can be used as an anti-biofilm agent to prevent biofilm formation of S. aureus and thus biofilm-related infections.

scholarly journals In vitro antifungal susceptibility of candidemia agents and detection of their biofilm production by two different methods

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Yasemin Oz ◽  
Iman Qoraan ◽  
Egemen Gokbolat
Keyword(s):  
Production Rate ◽  
Biofilm Formation ◽  
Colorimetric Assay ◽  
Formation Rate ◽  
Antifungal Susceptibility ◽  
Reference Method ◽  
Antifungal Resistance ◽  
Detection Methods ◽  
Biofilm Production

Candida bloodstream infections are a significant cause of morbidity and mortality in hospitalized patients. The most important contribution of biofilm is the higher antifungal resistance than planktonic cells. We aimed to investigate the biofilm formation rate and antifungal susceptibility characteristics of our bloodstream isolates, and evaluate two different biofilm detection methods. A total of 200 bloodstream Candida isolates were included. The biofilms were formed on 96-well microtiter plates and measured by spectrophotometric percent transmittance and 2,3-bis(2- methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium- 5-carboxanilide colorimetric assay. In addition antifungal susceptibilities of these isolates were evaluated against caspofungin, anidulafungin and amphotericin B by reference method. Biofilm production rate was considerably high among our bloodstream isolates. The most important biofilm producer species was C. tropicalis; C. glabrata had the lowest biofilm production rate. The consistency rate between biofilm detection methods was 66%. Remarkable antifungal resistance was not observed among our isolates in general. In conclusion, biofilm production in Candida species is an important virulence factor, and its rate is considerably high in bloodstream isolates. At present, a standardized method has not been established to detect the biofilm formation.

scholarly journals Biofilm formation by clinically isolated Staphylococcus Aureus from India

2018 ◽  
Vol 12 (12) ◽  
pp. 1062-1066 ◽  
Author(s):  
Alasthimannahalli Gangadhara Triveni ◽  
Mendem Suresh Kumar ◽  
Chavadi Manjunath ◽  
Channappa T Shivannavar ◽  
Subhaschandra M Gaddad
Keyword(s):  
Staphylococcus Aureus ◽  
Congo Red ◽  
Biofilm Formation ◽  
Quantitative Estimation ◽  
Microtiter Plate ◽  
Clinical Samples ◽  
Specific Method ◽  
Qualitative And Quantitative ◽  
Etiological Agents

Introduction: Staphylococcal biofilms are prominent cause for acute and chronic infection both in hospital and community settings across the world. Current study explores biofilm formation by Staphylococcus aureus isolates from clinical samples by different methods. Methodology: Standard techniques used for the characterization of S.aureus. Qualitative and quantitative biofilm formation was assessed by Congo red Agar, Tube and Microtiter plate methods. Results: A total of 188 clinical isolates of S.aureus were screened for biofilm formation and 72 (38.29%) of them were found to be biofilm producers, 34 (18.08%) strong, 38 (20.21%) moderate. The remaining 116 (61.7%) were weak/ non biofilm producers. Maximum biofilm formers were recorded in pus samples (39.06%), followed by isolates from blood (38.23%) and urine (34.61%). Statistical analysis for the formation of biofilm indicated that Microtiter plate method is the most sensitive and specific method for screening biofilm production. Conclusions: Biofilm formation is one of the influential virulence factor in staphylococcal pathogenesis and persistence. Microtiter plate and Congo red agar remain as reliable methods for the qualitative and quantitative estimation of biofilm formation. Monitoring of biofilm formation in various etiological agents will help in determining the severity of infection.

scholarly journals Adaptation of Congo Red Agar Method and Microtiter Plate Assay to Study Biofilm Formation in Streptomyces

2021 ◽  
Vol 18 (1) ◽  
pp. 113-123
Author(s):  
Rabha EL othmany ◽  
Hafida Zahir ◽  
Chorouk Zanane ◽  
Doha Mazigh ◽  
Mostafa Ellouali ◽  
...  
Keyword(s):  
Crystal Violet ◽  
Congo Red ◽  
Biofilm Formation ◽  
Staining Method ◽  
Microtiter Plate ◽  
Alcoholic Solution ◽  
Bioactive Molecules ◽  
Microtiter Plate Assay ◽  
Agar Method ◽  
Quantitative And Qualitative Methods

Streptomyces has many advantages for exploration in biotechnological applications because of their ability to elaborate a multitude of bioactive molecules and secondary metabolites. Despite the importance of this genus in biotechnology, biofilm formation in Streptomyces is under-investigated. The objective of this research is to adapt two assays for the assessment of biofilm formation in Streptomyces. In the present investigation, we assess and follow biofilm formation in eight Streptomyces strains using quantitative and qualitative methods. The quantitative study based on a staining of the retained biomass in the microtiter plate with crystal violet “5%” and destaining using ethanol/acetone mixture, the concentration of crystal violet in the alcoholic solution reflect the intensity of the attached biofilm. On the other hand, the qualitative one consists of using modified freeman’s method a modified congo red agar method based on the color of colonies. Quantification of biomass by crystal violet staining method confirmed that Streptomyces bellus A43 and Streptomyces bellus A61 are biofilm-forming and this ability increase with the period of incubation. Our results showed that sixStreptomyces strains arenon-slime producing/non-biofilm forming. Two Streptomyces strains are slime producing/biofilm forming; this character vanishes at five days. Further research on genes responsible for biofilm formation in Streptomyces is highly recommended for better understanding of the phenomenon.

scholarly journals Deletion of SarX decreases biofilm formation of Staphylococcus aureus in a polysaccharide intercellular adhesin (PIA) dependent manner

2020 ◽  
Author(s):  
Zhihao Hao ◽  
Yinjuan Guo ◽  
Lulin Rao ◽  
Jingyi Yu ◽  
Qing Zhan ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Mutant Strain ◽  
Biofilm Formation ◽  
Transcriptional Activity ◽  
Extracellular Proteins ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Biofilm Production ◽  
Fold Reduction ◽  
Operon Expression

Abstract Background: Biofilm formation by Staphylococcus aureus is an important virulence determinant mediated by the polysaccharide intercellular adhere (PIA) encoded by ica operon or by surface and extracellular proteins. Previous studies have shown that S. epidermidis SarX protein regulated the transcriptional activity of the agr and ica loci and controled the biofilm phenotype, primarily by regulating icaADBC transcription and PIA production.Results: In this study, our results indicated that biofilm formation and detachment of S. aureus were significantly decreased in the sarX mutant strain. sarX mutant in S. aureus biofilm formation was related to the production of PIA and not to that of eDNA. RT-PCR results showed that deletion of sarX was associated with a 1.8-fold reduction in spa transcription, which was complemented by sarX. Expression of Spa protein was decreased in sarX mutant strain.Conclusions: sarX promoted biofilm production of S. aureus may mediated primarily through increasing ica operon expression and PIA production. Deletion of sarX was associated with reduction in spa transcription.

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