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 Sub-Inhibitory Concentrations of Rifampicin Strongly Stimulated Biofilm Production in S. aureus

2017 ◽  
Vol 11 (1) ◽  
pp. 142-151 ◽  
Author(s):  
Agostinho Alves Lima-e-Silva ◽  
Renato Geraldo Silva-Filho ◽  
Henry Marcel Zalona Fernandes ◽  
Carmen Soares Meirelles Saramago ◽  
Alice Slotfeldt Viana ◽  
...  
Keyword(s):  
Medical Devices ◽  
Congo Red ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Proteinase K ◽  
Implantable Medical Devices ◽  
Sodium Metaperiodate ◽  
Microtiter Plate Assay ◽  
Biofilm Production ◽  
Very High

Background and Objectives:Staphylococcus aureusis an important pathogen and a frequent cause of infections associated with biofilm production in implantable medical devices. Biofilm production can be induced by sub-inhibitory concentrations (sub-MICs) of certain antibiotics, but few studies have researched this occurrence inS. aureus. In this study, we investigated the effect of sub-MICs of rifampicin and minocycline on biofilm production by five clinical and five non-clinicalS. aureusisolates.Methods:Microtiter Plate assay and Congo Red Agar Test were used to analyze the biofilm production. The biofilm composition was evaluated by the detachment assay with sodium metaperiodate and proteinase K.Results:Rifampicin sub-MICs induced very high biofilm formation in seven isolates that were non-producers in Tryptic Soy Broth. In one producer isolate, the biofilm formation level was not affected by sub-MICs of this drug. Sub-MICs of minocycline did not induce biofilm production in all isolates tested and in two producer isolates, instead, MIC/2 and MIC/4 inhibited biofilm production. The results of the drugs in combination were similar to those with rifampicin alone. The biofilm matrix was identified as polysaccharide, except for one producer isolate, classified as proteinaceous. Polysaccharide biofilm producer isolates, when grown on Congo Red Agar without sucrose, but with sub-MICs of rifampicin, showed results in agreement with those obtained in Microtiter Plate Test.Conclusion:The high biofilm production induced by sub-MICs of rifampicin has potential clinical relevance, because this is one of the drugs commonly used in the impregnation of catheters. In addition, it is used adjunctively to treat certainS. aureusinfections.

scholarly journals Microtiter Plate Assay for Assessment of Listeria monocytogenes Biofilm Formation

2002 ◽  
Vol 68 (6) ◽  
pp. 2950-2958 ◽  
Author(s):  
D. Djordjevic ◽  
M. Wiedmann ◽  
L. A. McLandsborough
Keyword(s):  
Listeria Monocytogenes ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Cellular Growth ◽  
Epifluorescence Microscopy ◽  
Simple Method ◽  
Microtiter Plate Assay ◽  
Plate Assay ◽  
Biofilm Production

ABSTRACT Listeria monocytogenes has the ability to form biofilms on food-processing surfaces, potentially leading to food product contamination. The objective of this research was to standardize a polyvinyl chloride (PVC) microtiter plate assay to compare the ability of L. monocytogenes strains to form biofilms. A total of 31 coded L. monocytogenes strains were grown in defined medium (modified Welshimer's broth) at 32°C for 20 and 40 h in PVC microtiter plate wells. Biofilm formation was indirectly assessed by staining with 1% crystal violet and measuring crystal violet absorbance, using destaining solution. Cellular growth rates and final cell densities did not correlate with biofilm formation, indicating that differences in biofilm formation under the same environmental conditions were not due to growth rate differences. The mean biofilm production of lineage I strains was significantly greater than that observed for lineage II and lineage III strains. The results from the standardized microtiter plate biofilm assay were also compared to biofilm formation on PVC and stainless steel as assayed by quantitative epifluorescence microscopy. Results showed similar trends for the microscopic and microtiter plate assays, indicating that the PVC microtiter plate assay can be used as a rapid, simple method to screen for differences in biofilm production between strains or growth conditions prior to performing labor-intensive microscopic analyses.

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 Biofilm production and its effects on virulence of MRSA: a study in tertiary care hospital, Delhi

2020 ◽  
Vol 7 (9) ◽  
pp. 3518
Author(s):  
Amir Khan ◽  
Rachna Tewari ◽  
Neetusree . ◽  
Mridu Dudeja
Keyword(s):  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Automated System ◽  
Assay Method ◽  
Common Source ◽  
Care Hospital ◽  
Microtiter Plate Assay ◽  
Biofilm Production ◽  
Antibiotic Susceptibility Test ◽  
Mrsa Strains

Background: The aim of this study was to study the prevalence of biofilm formation in MRSA and its effect on virulence and the antimicrobial resistance pattern on MRSA strains from different clinical samples.Methods: A total of 221 isolates of S. aureus isolates were selected from various clinical specimens. Prevalence was estimated according to age, sex, and location.  The antibiotic susceptibility test was conducted according to the guidelines of CLSI by the VITEK 2 automated system. 113 strains were identified as MRSA by cefoxitin disc methods which were then subjected to Microtiter plate assay method to confirm phenotypic biofilm formation.Results: 51.13% isolates were resistant to methicillin, and 48.86% isolates were methicillin sensitive. The most common source of MRSA isolation was blood. MRSA isolates were mostly isolated from male. 33.63% MRSA and 19.44% MSSA isolates were strong biofilm producers while 12.38% MRSA and 14.81% were low biofilm producers. The resistance for commonly used antibiotics like benzyle penicillin, ciprofloxacin, cotrimoxazole, and erythromycin was more in MRSA strains and MIC was higher in biofilm producers.Conclusion: Statistical difference was observed between MSSA and MRSA regarding biofilm formation and antimicrobial resistance. A Biofilm producer shows resistance to many antibiotics and also make host immunity in effective. In hospitals Biofilm production should be checked regularly before giving treatment. And research should be done to find out other effective drugs to eradicate biofilms. 

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 Correlation of biofilm formation and antibiotic resistance among clinical and soil isolates of Acinetobacter baumannii in Iraq

2019 ◽  
pp. 1-10 ◽  
Author(s):  
Pakhshan A. Hassan ◽  
Adel K. Khider
Keyword(s):  
Acinetobacter Baumannii ◽  
Congo Red ◽  
Biofilm Formation ◽  
Opportunistic Pathogen ◽  
Microtiter Plate ◽  
Test Tube ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Disk Diffusion Method

Acinetobacter baumannii is an opportunistic pathogen that is reported as a major cause of nosocomial infections. The aim of this study was to investigate the biofilm formation by A. baumannii clinical and soil isolates, to display their susceptibility to 11 antibiotics and to study a possible relationship between formation of biofilm and multidrug resistance. During 8 months period, from June 2016 to January 2017, a total of 52 clinical and 22 soil isolates of A. baumannii were collected and identified through conventional phenotypic, chromo agar, biochemical tests, API 20E system, and confirmed genotypically by PCR for blaOXA-51-like gene. Antibiotic susceptibility of isolates was determined by standard disk diffusion method according to Clinical and Laboratory Standard Institute. The biofilm formation was studied using Congo red agar, test tube, and microtiter plate methods. The clinical isolates were 100% resistance to ciprofloxacin, ceftazidime, piperacillin, 96.15% to gentamicin, 96.15% to imipenem, 92.31% to meropenem, and 78.85% to amikacin. The soil A. baumannii isolates were 100% sensitive to imipenem, meropenem, and gentamicin, and 90.1% to ciprofloxacin. All A. baumannii isolates (clinical and soil) were susceptible to polymyxin B. The percentage of biofilm formation in Congo red agar, test tube, and microtiter plate assays was 10.81%, 63.51%, and 86.48%, respectively. More robust biofilm former population was mainly among non-MDR isolates. Isolates with a higher level of resistance tended to form weaker biofilms. The soil isolates exhibited less resistance to antibiotics than clinical isolates. However, the soil isolates produce stronger biofilms than clinical isolates.

CHARACTERIZATION OF BACTERIAL ADHESION AND BIOFILM FORMATION ABILITY OF LIVESTOCK-ASSOCIATED METHICILLIN-RESISTANTSTAPHYLOCOCCUS AUREUSISOLATED FROM SWINE AND SLAUGHTERHOUSE WASTEWATERS

2014 ◽  
Vol 40 (02) ◽  
pp. 101-107
Author(s):  
Min-Tao Wan ◽  
Chin-Cheng Chou
Keyword(s):  
Bacterial Adhesion ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Ecological Niches ◽  
Microtiter Plate Assay ◽  
Environmental Group ◽  
Biofilm Production ◽  
The Difference ◽  
Wastewater Samples

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) ST9 has emerged as a potential zoonotic pathogen for humans and animals. Bacterial adhesion factors and biofilms mediate host colonization and infection of MRSA. This study investigated the dynamics of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs), biofilm formation gene (intercellular adhesion [ica]), and biofilm expression in MRSA from the nasal samples of asymptomatic pigs (the nasal group, n = 147) and swine slaughterhouse wastewater samples (the environmental group, n = 86). Biofilm formation was quantified by microtiter plate assay. The most prevalent MSCRAMM profile was clfA-clfB-spa-eno-ebps-fib and more than 70% of the LA-MRSA ST9 isolates harbored the biofilm formation gene. Environmental MRSA harbored lower levels of the ica locus and MSCRAMMs (clfA and fib) than did the nasal group, suggesting possible gene loss. Biofilm production in the nasal group was higher than in the environmental group, indicating the difference in biofilm formation in MRSA isolates from different ecological niches. The higher prevalence of MSCRAMMs, biofilm formation gene, and biofilm production in LA-MRSA ST9 may enhance the persistence and infectivity of MRSA in the swine population and present a threat to the health of livestock as well as farm workers.

scholarly journals Study on biofilm-forming properties of clinical isolates of Staphylococcus aureus

2011 ◽  
Vol 6 (05) ◽  
pp. 403-409 ◽  
Author(s):  
Yasmeen Taj ◽  
Farhan Essa ◽  
Faisal Aziz ◽  
Shahana Urooj Kazmi
Keyword(s):  
Electron Microscopy ◽  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Assay Method ◽  
Scanning Electron Micrographs ◽  
Clinical Specimens ◽  
Biofilm Production ◽  
Important Virulence Factor ◽  
Air Liquid Interface ◽  
Scanning Electron

Introduction: The purpose of this study was to observe the formation of biofilm, an important virulence factor, by isolates of Staphylococcus aureus (S. aureus) in Pakistan by different conventional methods and through electron microscopy. Methodology: We screened 115 strains of S. aureus isolated from different clinical specimens by tube method (TM), air-liquid interface coverslip assay method, Congo red agar (CRA) method, and scanning electron microscopy (SEM). Results: Out of 115 S. aureus isolates, 63 (54.78%) showed biofilm formation by tube method. Biofilm forming bacteria were further categorized as high producers (n = 23, 20%) and moderate producers (n = 40, 34.78%). TM coordinated well with the coverslip assay for strong biofilm-producing strains in 19 (16.5%) isolates. By coverslip method, weak producers were difficult to differentiate from biofilm negative isolates. Screening on CRA showed biofilm formation only in four (3.47%) strains. Scanning electron micrographs showed the biofilm-forming strains of S. aureus arranged in a matrix on the propylene surface and correlated well with the TM. Conclusion: Biofilm production is a marker of virulence for clinically relevant staphylococcal infections. It can be studied by various methods but screening on CRA is not recommended for investigation of biofilm formation in Staphylococcus aureus. Electron micrograph images correlate well with the biofilm production as observed by TM. 

scholarly journals Determination of Biofilm Formation between Different Strains of Propionibacterium acnes Using Biofilm Assay

2019 ◽  
Vol 4 (02) ◽  
Author(s):  
Yusufu, W. N. ◽  
Suleiman, H. O. ◽  
Akwa, V. Y. ◽  
David, D. L. ◽  
Taiga, A.
Keyword(s):  
Biofilm Formation ◽  
Propionibacterium Acnes ◽  
Acne Vulgaris ◽  
Microtiter Plate ◽  
Deep Tissue ◽  
Microtiter Plate Assay ◽  
Biofilm Production ◽  
Culture Isolate ◽  
Different Strains

Propionibacterium acnes (P. acnes) a member of the normal flora of the skin has constantly been associated with deep tissue infections especially during medical processes. P. acnes have been isolated in deep tissues and are believed to be an aetiological agent in these infections, contributing to the progression of some of these diseases. The biofilm formation ability between different strains of P. acnes was determined. Ten (10) P. acnes clinical isolates were considered, two (2) from acne vulgaris and eight (8) [two (2) per recA types 1A1, 1B, II and III] from lumber herniation tissues. Semi quantitative biofilm analysis using the microtiter plate assay was used with some modification. The semi quantitative biofilm assay was done in triplicates. The result obtained from the biofilm triplicates from 4 days’ incubation using 3 days’ culture showed that isolates 17(IB), 82(IB) and 55(II) showed very high biofilm production for 2 replicates which implies that they are real biofilm producing isolates. Using overnight cultures, higher biofilm production was witnessed with isolates 1(III), Lesion 7 and 84 (IA1) being the highest biofilm producers. Although with 3 days’ culture, isolate 1(III) could easily be discarded as a-non biofilm producer, while lesion 7 and 84 (IA) has been associated to biofilm formation in 3 days’ culture. The production of biofilms by isolates supports the theory that the ability of P. acnes to form biofilms enables it to attach to medical implants hence causing deep tissue infections.

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