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1
Normanita, Rina. "Validity Of Congo Red Agar And Modified Congo Red Agar To Detect Biofilm Of Enterococcus Faecalis." Saintika Medika 16, no. 1 (2020): 55. http://dx.doi.org/10.22219/sm.vol16.smumm1.11064.
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Purpose: Enterococcus faecalis causes nosocomial infections such as bacteremia, urinary tract infections, intra-abdominal infections, and endocarditis. These infection is associated with biofilm and intrinsically resistant to many antibiotics. This study aims to determine the validity of the CRA and MCRA for detecting biofilms of Enterococcus faecalis Method: This is a laboratory observational study with 30 sample of Enterococcus faecalis. We performed biofilm examination for Enterococcus faecalis by using Congo red Agar, Modified Congo red Agar and Microtitter Plate Assay as gold standard. Result: Both MCRA and CRA were compared MPA as a gold standard was obtained p value is 0.309 (p> 0.05), with a Kappa agreement coefficient is 0.067, which indicates there is no significant agreement to detect biofilm of Enterococcus faecalis. MCRA and CRA have almost no compatibility with MPA for biofilm forming of Enterococcus faecalis. Conclusion: Both MCRA and CRA has a very high sensitivity (100%), but the specificity is very low 6.67% for detecting the biofilms of Enterococcus faecalis. MCRA and CRA can not determine negativity well and it have a high false positive rate, so to increase specificity of biofilm forming, we must combine these method with the others. Keywords: Biofilm, Enterococcus faecalis, CRA, MCRA, MPA.
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Abdel Halim, Rania M., Nevine N. Kassem, and Basma S. Mahmoud. "Detection of Biofilm Producing Staphylococci among Different Clinical Isolates and Its Relation to Methicillin Susceptibility." Open Access Macedonian Journal of Medical Sciences 6, no. 8 (2018): 1335–41. http://dx.doi.org/10.3889/oamjms.2018.246.
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AIMS: To evaluate three in vitro phenotypic methods; tissue culture plate, tube method, and Congo red agar for detection of biofilm formation in staphylococci and assess the relation of biofilm formation with methicillin resistance and anti-microbial resistance.
 METHODS: The study included 150 staphylococcal isolates. Biofilm detection in staphylococci was performed using tissue culture plate, tube method, and Congo red agar.
 RESULTS: Tissue culture plate, tube method, and Congo red agar detected 74%, 42.7%, and 1.3% biofilm producing staphylococci respectively. S. aureus isolates were more common biofilm producers (53.2%) than CONS (46.8%). Biofilm production in CONS species was highest in S. hemolyticus (57.7%). Tube method was 51.4% sensitive, 82.1% specific. As for Congo red agar, sensitivity was very low (0.9%), but specificity was 97.4%. Biofilm producers were mostly; isolated from blood specimens (82.6%) and detected in methicillin-resistant strains 96/111 (86.5%). They were resistant to most antibiotics except vancomycin and linezolid.
 CONCLUSIONS: Tissue culture plate is a more quantitative and reliable method for detection of biofilm producing staphylococci compared to tube method and Congo red agar. Hence, it can still be used as a screening method for biofilm detection. Vancomycin and Linezolid are the most sensitive antibiotics among biofilm producing staphylococci.
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Al-yozbakee, Zinah Makki, and Khalid Omar Mohammad. "EVALUATION OF MODIFIED CONGO RED AGAR FOR DETECTION OF BIOFILM PRODUCING MDR KLEBSIELLA PNEUMONAIE CLINICAL ISOLATES." European Journal of Medical Genetics and Clinical Biology 1, no. 8 (2024): 89–101. https://doi.org/10.61796/jmgcb.v1i8.775.
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The notoriety of Multiple Drug Resistant (MDR) K. pneumoniae extends beyond nosocomial or community acquired infections, since it is a major cause of biofilm-related infections. The utilization of a very specific or distinct media, Congo Red Agar (CRA), for detection of biofilm, has significant drawbacks, including inconsistencies in the creation of black pigment. In this study, an assessment of Modified Congo Red Agar (MCRA), for detection of biofilm, was performed on 23 multidrug-resistant (MDR) K. pneumoniae strains obtained from various clinical samples. The aim of this study was to assess the effectiveness and dependability of Modified Congo Red Agar (MCRA) as an alternative medium for studying biofilm production to the non modified Congo Red Agar. Also to examine the prevalence of the fimH gene, which is necessary for biofilm formation among these isolates. Lastly to determine the relationship between antibiotic resistance, the presence of the fimH gene, and the ability to produce biofilm among K. pneumonaie strains. These strains were diagnosed according to their specific bacteriological characteristics. After assessing antibiotic susceptibility, all MDR K. pneumoniae isolates exhibited the presence of the fimH gene using the PCR method. On CRA 20 isolates were strong biofilm producer as they appeared as black colored colonies, two were moderate biofilm producer (pink colored) and only 1 was non biofilm producer ( red colored) colony. On MCR the same results for biofilm production to CRA, except for one colony differed as it appeared red (non biofilm producer) on MCRA and pink (moderate biofilm producer) on CRA. However, the growth of 75% blackness pigment of MDR K. pneumoniae strains on the CRA decreased over time. The phenotypic pigmentation on CRA was enhanced by modifying the contents of the agar that led to the persistent development of a highly concentrated black pigment in isolates containing the fimH gene for 2 to 4 days, with no drop in pigmentation seen over time. The change of the agar ingredient enabled the stable synthesis of black pigment and also resulted in a reduction in the cost of agar preparation. Key Words: Biofilm, Congo Red Agar, fimH gene, Modified Congo Red Agar, MDR K. pneumonaie.
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Sabaa, Hanaa Salih. "Elimination of Biofilm Produced by Klebsiella Pneumonia Using Strontium Radioactive Sources (Sr90)." NeuroQuantology 20, no. 2 (2022): 76–81. http://dx.doi.org/10.14704/nq.2022.20.2.nq22028.
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The aim of this study was conducted to elimination biofilm production from K. pneumonia by Sr90 radiosources inorder to removal biofilm that contained with K. pneumonia infection by beta ray. Study design and study populations of 100 isolates of K. pneumonia that identified by Vitek2GN, results of identification for all isolates approve are K. pneumonia. Study production of biofilm achieved by Congo red agar test, positive results by formation black colony on Congo red agar and negative results with red colony on Congo red agar. Influence Strontium Sr90 on K. pneumonia with dose D*(1hr.) 0.73251*10-2 msV; D*(2hr.) 1.46502*10-2 msV; D*(3hr.) 2.19753*10-2 msV; D*(4hr.) 2.93004*10-2 msV in different hours percentage of killing for this bacteria reach to 90% with control 300 colony, elimination of biofilm production from this bacteria by all doses in different time with formation pink color of colony compared with control formation black colony on Congo red agar, this results of affect by Strontium cause lost production biofilm by turn off into pink color when exposed to different doses of beta rays emitted from Strontium compared with control with black color.
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Karreman, Robert J., and George G. Lindsey. "Modulation of Congo-red-induced aberrations in the yeast Saccharomyces cerevisiae by the general stress response protein Hsp12p." Canadian Journal of Microbiology 53, no. 11 (2007): 1203–10. http://dx.doi.org/10.1139/w07-090.
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Previous studies have shown that in Saccharomyces cerevisiae HSP12, which codes for the small cell wall heat shock protein Hsp12p, was induced upon exposure to cell-wall-damaging agents such as Congo red. Here, we demonstrate that Hsp12p decreases the interaction between Congo red and chitin. A Δhsp12 mutant strain displayed decreased viability, increased aggregation and sedimentation, as well as an altered morphology when grown in the presence of Congo red dye. The presence of Hsp12p was also necessary for the Congo-red-mediated invasion of agar plates.
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Worsham, Patricia L., and Michele R. Sowers. "Isolation of an asporogenic (spoOA) protective antigen-producing strain of Bacillus anthracis." Canadian Journal of Microbiology 45, no. 1 (1999): 1–8. http://dx.doi.org/10.1139/w98-108.
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We found that Congo red agar allows identification of sporulation-deficient Bacillus anthracis. Using Congo red agar, we isolated an asporogenic derivative of the protective antigen-producing strain B. anthracis deltaSterne-1(pPA102). Polymerase chain reaction and Southern hybridization analyses of DNA from the asporogenic mutant revealed that a deletion was present in spoOA, an essential gene for the initiation of sporulation. The deletion also encompassed the spoIVB homologue and a portion of the recN homologue. The avirulent spoOA strain deltaSterne-1(pPA102)CR4 is suitable for the safe production of protective antigen without endospore contamination of the vaccine production facility.Key words: Bacillus anthracis, protective antigen, spoOA, vaccine, Congo red.
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Fiadh, Hadeel M. "Detection of biofilm formation among the mastitis isolates of Staphylococci by evaluation of three different screening methods." Kufa Journal For Veterinary Medical Sciences 2, no. 2 (2011): 107–15. http://dx.doi.org/10.36326/kjvs/2011/v2i23892.
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Internalization and biofilm formation is an important step in staphylococcal mastitis pathogenesis . Twenty five staphylococcal isolates of coagulase negative and coagulase positive were collected from clinical mastitis in cows, and biofilm production were studied for these isolate by three different methods. Tube method , tissue culture plate and Congo red agar method. The first two methods were used to study effect of glucose addition as source of polysaccharides that important in biofilm formation. The results showed significant increase in biofilm formation by addition of glucose in staphylococci coagulase negative diagnosed by tissue culture plate method. Congo red agar method results showed increase number of positive isolates by increasing incubation time. The results showed that tissue culture plate method (Spectrophotometric method) was the best method for the diagnosis of biofilm formation in comparison with both tube method and Congo red agar method.
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Deighton, M. A., J. Capstick, and R. Borland. "A study of phenotypic variation ofStaphylococcus epidermidisusing Congo red agar." Epidemiology and Infection 109, no. 3 (1992): 423–32. http://dx.doi.org/10.1017/s095026880005041x.
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SUMMARYThis study examines a series of phenotypic variants ofStaphylococcus epidermidisthat were generated from a pair of parent variants, isolated from valvular tissue of a patient with prosthetic valve endocarditis. The variants were initially classified by examining their colonial morphology on Congo red agar. In addition to differences in Congo red binding and colonial morphology, they differed in the expression of several surface components and enzymes. Despite these phenotypic differences, all variants had the same restriction endonuclease profile of plasmid DNA. Examination of a collection of clinical isolates demonstrated that phenotypic variation is a common property ofS. epidermidis. The ability to express different combinations of surface components and enzymes could contribute to the virulence ofS. epidermidisstrains by enabling these organisms to colonize a range of diverse environments.
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Furtuna, Dewi Klarita, Kartuti Debora, and Eddy Bagus Warsito. "Comparison of Microbiological Examination by Test Tube and Congo Red Agar Methods to Detect Biofilm Production on Clinical Isolates." Folia Medica Indonesiana 54, no. 1 (2018): 22. http://dx.doi.org/10.20473/fmi.v54i1.8047.
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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.
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Jabir, Dhuha. "Evaluation of the ability of some bacterial species isolated from UTIto form bio film." Al-Qadisiyah Journal of Pure Science 27, no. 1 (2022): 8–14. http://dx.doi.org/10.29350/qjps.2022.27.1.1492.
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Urinary tract infection is one of the most serious infectious disease in the world , This study aimed to isolate and diagnose the bacteria that cause UTI and then evaluate its ability to form a biofilm. 100 urine samples were collected for a group of patients attending Al-Diwaniyah Teaching Hospital, who were confirmed to have UTI at different ages. The results showed the presence of bacteria. Escherichia coli and proteus spp in all samples(100%), while Staphylococcus aureus was found in 87% of samples , Enterobacter spp was isolated by 67% and Pseudomonas aeruginosa by 65%. Klebsiella pneumonia at 50 % and Staphylococcus epidermidis and Micrococcus spp each with 30%. Regarding the ability of the isolated bacteria to form biofilm, two methods used Congo-red agar and tube method and the results were as follows 64% for E.coli, K pneumonia with 66.6% while S. aureus showed the ability of formation with 41.6% and negative results for the rest by using tube method while Congo-red agar method results were 25% for 50% for E.coli and S. aureus. The study concluded that Congo red agar method is easy to perform and interpret, while the tube method is highly sensitive and specific.
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Mehta, Mrs. Ishwari Nitesh, and Dr. Shilpa M. Gharat. "Biodegradation of Direct Azo Dye-Congo Red Using the Bacterial Isolates from Waste Landfill in Khaira Phata - Boisar, Maharashtra." International Journal of Advance and Applied Research 6, no. 25 (2025): 184–87. https://doi.org/10.5281/zenodo.15294806.
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<strong>Abstract: </strong> Congo red- Direct Azo dye one of the synthetic dye used in textile industry for dyeing cotton. Its discharge in the water bodies as textile effluents has affected the flora and fauna of the ecosystem and also entered in the agricultural land causing many health hazards in humans. The present study emphasized on the degradation of the Direct Azo dye- Congo red by using the bacterial isolates from the waste land fill areas of Khaira-phata region of Boisar, Maharashtra. In the study carried out, the azo dye degrading organisms were isolated and selected by the crowded plate technique, the selective plates contained the Nutrient agar containing 20 ppm of the Congo red. The colonies showing the zone of clearance around them were selected and further identified by microscopic, cultural and biochemical characteristics was found to be Gram positive cocci. 100 ppm of Congo red was used to check the efficiency of decolourization. Efficient decolourization of 100 ppm of Congo red was obtained after 120 hours and was found to be 77.77 % under static conditions at room temperature by the bacterial isolate .
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Mishra, Durgesh, Arpita Shrivastava, Neeraj Shrivastava, Rajeev Ranjan, and Ankush Kiran Niranjan. "Study of Antimicrobial and Antibiofilm Effect of Essential Oils on Biofilm Producing Staphylococcus aureus Isolated from Mastitis Milk of Bovines." Indian Journal of Veterinary Sciences and Biotechnology 21, no. 2 (2025): 37–42. https://doi.org/10.48165/ijvsbt.21.2.08.
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The present work was undertaken to study antimicrobial and antibiofilm effect of essential oils on the biofilm producing Staphylococcus aureus isolated from the bovine mastitis milk. Out of 164 dairy cattle and buffalo milk samples screened, 24 samples were found positive for mastitis by CMT. Among these, 17 S. aureus isolates were identified and characterized phenotypically by standard biochemical tests. The biofilm formation ability of the isolates was studied by Congo red agar, Light microscopy method and Microtiter plate (Crystal Violet) method. The antimicrobial activity and antibiofilm effect of essential oils - Garlic and Cinnamon oil - on biofilm positive S. aureus were studied by agar well diffusion method and Microtiter plate method, respectively. Cefoperazone was used as standard control drug. Out of the total 17 S. aureus strains,70% were positive on Congo red agar, 52% by Light microscopy and 100% on Microtiter plate method. By the Microtiter plate method, 4 strains were strong biofilm producers and 13 were weak biofilm producers. Cinnamon oil showed better antibiofilm activity in all the concentration used (1%, 2%, 3%). Garlic oil showed good antimicrobial and antibiofilm activity at 3% concentration.It was concluded that biofilm producing Staphylococcus aureus was isolated from bovine mastitis milk samples, and sensitivity of Congo red agar, Light microscopy and Microtiter plate methods was good. Micro titre plate showed highest sensitivity for detection of biofilm production. Garlic oil and Cinnamon oil showed antimicrobial and antibiofilm producing abilityagainst S. aureus.
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Uzuegbunam, Ngozi V., Faruk A. Umar, and Bassey Enya Bassey. "Biofilm Detection on Catheter Associated Uropathogenic Bacteriuria among Fistula Patients Attending National Obstetric Fistula Centre Ningi, Bauchi State, Nigeria." European Journal of Medical and Health Sciences 3, no. 2 (2021): 180–84. http://dx.doi.org/10.24018/ejmed.2021.3.2.803.
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Background: Biofilm production caused by bacteria plays a vital role in catheter associated urinary tract infection (UTI) or bacteriuria being responsible for persistence and recurrent infection. Biofilms forming bacteria are difficult to eradicate due to antimicrobial resistance to the commonly used antibiotic. Biofilms are currently estimated to be responsible for over 65% of nosocomial infections and 80% of microbial infections. This study aimed to perform biofilm detection on uropathogenic bacterial isolates among fistula patients attending National Obstetric Fistula centre Ningi and investigate the antimicrobial susceptibility pattern.
 Methods: A total of 217 strains of significant bacteriuria were isolated from vesico vaginal fistula (VVF) patients. A cross sectional study was conducted at the hospital. The urine samples were collected and cultured on CLED and blood agar media while confirmation was done using their biochemical reaction. The detection of biofilms formation on the isolates was performed using tube adherence and Congo red agar method. Antimicrobial susceptibility testing was carried out by disc diffusion method on Muller Hinton agar.
 Results: Out of 217 significant bacteriuria isolated, 38 strains produced biofilms;28 strains tested positive on tube adherence method while 15 strains were positive on Congo red agar method. Bacteria that produced biofim showed multiple drug resistance compared to the platonic bacterial cells. All the biofilm producers showed 100% resistant to septrin, ampiclox, gentamycin and amoxicillin. There was no significant value between tube adherence and Congo red agar method with P value > 0.05.
 Conclusion: Biofilm detection should form part of routine testing while antimicrobial susceptibility testing is paramount on better choice of antibiotic therapy for proper management to reduce economic lost, treatment failure and drug resistance.
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Türkyılmaz, Süheyla, and Seyhan Kaynarca. "The Slime Production by Yeasts Isolated from Subclinical Mastitic Cows." Acta Veterinaria Brno 79, no. 4 (2010): 581–86. http://dx.doi.org/10.2754/avb201079040581.
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The aim of this study was to isolate yeasts from subclinical mastitic cows and to investigate the slime production by the isolated yeasts. The material used in this study included 339 milk samples from 152 dairy cattle with subclinical mastitis.Milk wasplated onto blood agar, MacConkey agar and Sabouraud dextrose agar. Forty-onesamples (12.1% of total milk samples) were found positive for the yeast by API 20 C AUX identification system. The isolated yeasts were classified into four genera ofCandida, Trichosporon, CryptococcusandSaccharomyces.TheCandidaspecies were following:C. krusei,C. kefyr,C. guilliermondii,C. famata,C. rugosaandC. utulis. Other yeasts were identified asTrichosporon mucoides,T. asahii,Cryptococcus laurentii,C. neoformansandSaccharomyces cerevisiae. Slime production was tested on Congo red brain heart infusion agar and evaluated according to Congo red phenomenon. Fifteen (36.6%) strains were slime factor positive: seven wereC. krusei, fourC. kefyr, oneC. guilliermondii, oneC. famata, oneT. asahii, and oneC. laurentii. The results of the present study indicate that yeast mastitis is significant for causing economic losses and slime production is mostly found in non-albicans Candidaspecies. Therefore, non-albicans Candidaspecies should be examined for slime production.
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Abdelsalam, M., K. Nakanishi, K. Yonemura, T. Itami, S. C. Chen, and T. Yoshida. "Application of Congo Red agar for detection ofStreptococcus dysgalactiaeisolated from diseased fish." Journal of Applied Ichthyology 25, no. 4 (2009): 442–46. http://dx.doi.org/10.1111/j.1439-0426.2009.01226.x.
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Ngieng, Ngui Sing, Azham Zulkharnain, Hairul Azman Roslan, and Ahmad Husaini. "Decolourisation of Synthetic Dyes by Endophytic Fungal Flora Isolated from Senduduk Plant (Melastoma malabathricum)." ISRN Biotechnology 2013 (July 22, 2013): 1–7. http://dx.doi.org/10.5402/2013/260730.
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A total of twenty endophytic fungi successfully isolated from Melastoma malabathricum (Senduduk) were examined for their ability to decolourise azo dyes: Congo red, Orange G, and Methyl red and an anthraquinone dye, Remazol Brilliant Blue R. Initial screening on the glucose minimal media agar plates amended with 200 mg L−1 of each respective dye showed that only isolate MS8 was able to decolourise all of the four dyes. The isolate decolourised completely both the RBBR and Orange G in the agar medium within 8 days. Further quantitative analysis of the dye decolourisation by isolate MS8 in aqueous minimal medium showed that isolate MS8 was able to decolourise all the tested dyes at varying levels. Dye decolourisation by the isolate MS8 was determined to be 97% for RBBR, 33% for Orange G, 48% for Congo red, and 56% for Methyl red, respectively, within a period of 16 days. Molecular identification of the fungal isolate MS8 using primer ITS1 and ITS4 showed that isolate MS8 shared 99% sequence similarity with Marasmius cladophyllus, a Basidiomycete. The ability to decolourise different types of dyes by isolate MS8 thus suggested a possible application of this fungus in the decolourisation of dyestuff effluents.
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Mohammad, Khadija Ismail, and Alaa Hussein Taha. "Investigating the ability of bacteria isolated from various artificial respiratory systems to resist antibiotics and biofilms formation." Science Archives 03, no. 04 (2022): 263–67. http://dx.doi.org/10.47587/sa.2022.3404.
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Respiratory infections associated with ventilators represent a significant risk to individuals in intensive care units, particularly children under 3 years of age. 100 sample were collected Between October 2021 and February 2022, from Mosul hospitals (Ibn Al-Atheer Teaching Hospital, Ibn Sina Teaching Hospital, Al-Salam Hospital, and Al-Khansa Teaching Hospital) at ages 1 day to 3 years. A swab was taken from the water chamber of the artificial breathing devices, and the catheter head was collected, where the mucous material was cultured in three different mediums (blood agar, MacConkey agar, and Chocolate agar). 80 bacterial isolates were collected. The highest percentage was determined to be connected to Gr- bacteria, with 65 isolates 81.25%, and 15 isolates 18.75% being Gr+ bacteria. Using the Kirby-Bauer disc diffusion method, it was discovered that all isolates had great resistance to the antagonist’s ampicillin and ceftriaxone, but high sensitivity to both Imipenem and Meropenem. The tube method was found to be more efficient than the Congo red agar method in the detection of biofilms. Klebsiella pneumoniae, Stenotrophomonas maltophilia, Burkholderia cepacia group, and Pseudomonas mendocina were biofilm-forming in both ways by up to 100%. The current investigation found that the tube approach produced 100% biofilm, but the Congo red agar method produced 41.25% biofilm.
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Lal, M., H. Kaur, and LK Gupta. "Differentiation between Virulent and Avirulent Y.enterocolitica Isolates using Congo Red Magnesium Oxalate Agar." Indian Journal of Medical Microbiology 21, no. 3 (2003): 214. http://dx.doi.org/10.1016/s0255-0857(21)03081-4.
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Riley, G., and S. Toma. "Detection of pathogenic Yersinia enterocolitica by using congo red-magnesium oxalate agar medium." Journal of Clinical Microbiology 27, no. 1 (1989): 213–14. http://dx.doi.org/10.1128/jcm.27.1.213-214.1989.
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Reshma, A. Amala, S. Niren Andrew, P. Saradhai, and S. Sasikala. "In vitro ANALYSIS OF BIOFILM FORMATION AND ANTIBIOTIC RESISTANCE OF UROPATHOGENIC Escherichia coli." Applied Biological Research 26, no. 1 (2024): 43–49. http://dx.doi.org/10.48165/abr.2024.26.01.5.
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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.
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Chalop, Karam Khalid, and Ibtisam Fadhil Moussa. "Investigation Ability of Local Micrococcus Luteus to Produce Biofilm (Sustainable Exopolysaccharide) and Studying Its Optimum Production Conditions." IOP Conference Series: Earth and Environmental Science 1487, no. 1 (2025): 012131. https://doi.org/10.1088/1755-1315/1487/1/012131.
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Abstract The study aimed to detection the ability of local Micrococcus luteus ML1 to produce Biofilm (Sustainable exopolysaccharides) & determining some of the optimal conditions for Its production by studying six factors affecting the production(temperature, pH, inoculame volume, incubation time, sucrouse percentage, Shaking speed) in the laboratories of Iraqi Ministry of Agriculture / Department of Crop Protection. Congo red agar test & Elisa Technique designated to investigate the ability of isolates to produce biofilm, as the isolate proved its ability to produce exopolysaccharides as indicated by the colour change of the Congo red agar medium as a result of the formation of the biofilm in this environment, also the isolate show a moderate biofilm production in microtiterplates. The bacteria gave the highest production of the extracellular polysaccharides ma45trix at a temperature of 35°C, an incubation period of 168 hours, a pH of 7, a sucrose concentration of 15%, an inoculum volume of 6 ml, and a shaking speed of 75 rpm.
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Hawkins, Justin P., Barney A. Geddes, and Ivan J. Oresnik. "Common dyes used to determine bacterial polysaccharides on agar are affected by medium acidification." Canadian Journal of Microbiology 63, no. 6 (2017): 559–62. http://dx.doi.org/10.1139/cjm-2016-0743.
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In this work, we highlight effects of pH on bacterial phenotypes when using the bacteriological dyes Aniline blue, Congo red, and Calcofluor white to analyze polysaccharide production. A study of galactose catabolism in Sinorhizobium meliloti led to the isolation of a mutation in dgoK1, which was observed to overproduce exopolysaccharides when grown in the presence of galactose. When this mutant strain was spotted onto plates containing Aniline blue, Congo red, or Calcofluor white, the intensity of the associated staining was strikingly different from that of the wild type. Additionally, a Calcofluor dull phenotype was observed, suggesting production of a polysaccharide other than succinoglycan. Further investigation of this phenotype revealed that these results were dependent on medium acidification, as buffering at pH 6 had no effect on these phenotypes, while medium buffered at pH 6.5 resulted in a reversal of the phenotypes. Screening for mutants of the dgoK1 strain that were negative for the Aniline blue phenotype yielded a strain carrying a mutation in tkt2, which is annotated as a putative transketolase. Consistent with the plate phenotypes, when this mutant was grown in broth cultures, it did not acidify its growth medium. Overall, this work shows that caution should be exercised in evaluating polysaccharide phenotypes based strictly on the use of dyes.
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Retina Paul, Priyanka Saha, Kuhu Pal, Avi Kapoor, Arpita Sinha, and Joydip Roy. "Study of biofilm formation among uropathogens isolated from catheter-associated UTI patient from a tertiary care hospital." Asian Journal of Medical Sciences 15, no. 2 (2024): 126–32. http://dx.doi.org/10.3126/ajms.v15i2.59265.
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Background: Catheter-associated urinary tract infection (CAUTI) is a serious health threat and challenging infection. CAUTI accounts for up to 40% of all nosocomial infections. Biofilm provides a survival strategy to microorganisms and ultimately leads to re-infections and recurrence of urinary tract infections (UTI) despite a full course of antibiotics. Aims and Objectives: The present study was conducted to determine the prevalence of CAUTI in suspected UTI patients and prevalence of biofilm-forming uropathogens among CAUTI patients. Materials and Methods: The cross-sectional study was done over a period of 6 months among 95 catheterized CAUTI patients. Biofilm production among isolated uropathogens was tested by tissue culture plate, tube test, and congo red agar method. Isolates were identified as biofilm producer if they were tested positive by any one of the all four methods, and isolates were considered biofilm non-producer in consensus with all four methods. Results: In this study, the prevalence of CAUTI was 68.42%. Among 65 isolates most common uropathogen was 28 (43.07%) Escherichia coli. In this present study, the prevalence of biofilm-forming uropathogens was 58.46% (38). Tissue culture plate was the most sensitive (97.36%) method in detecting biofilm formation followed by modified congo red agar (82.21%), congo red agar (71.05%), and tube test (65.78%). Biofilm productions were significantly associated with female gender, diabetes, and prolonged catheterization. Conclusion: Indwelling urinary catheter acts as a nidus for biofilm formation among microorganisms. Duration of catheterization is inversely associated with UTI. Hence, the need for catheter removal should be assessed daily to prevent infection. Periodic surveillance should be done to detect biofilm formation where prolonged catheterization is inevitable.
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Pradhan, Roshani Nhuchhen, Surendra Kumar Madhup, and Shyam Prasad Pant. "Antibiogram and Biofilm Formation among Carbapenem Resistant Klebsiella pneumoniae." Tribhuvan University Journal of Microbiology 6 (December 7, 2019): 63–69. http://dx.doi.org/10.3126/tujm.v6i0.26586.
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Objectives: This cross-sectional study was designed to detect the carbapenemase producing K. pneumoniae along with biofilm producers from different clinical specimens and to compare antibiotic susceptibility pattern of biofilm producing carbapenem resistant Klebsiella pneumoniae and biofilm non-producing carbapenem resistant Klebsiella pneumoniae.
 Methods: A total of 1475 non-repetitive clinical samples were included on this study. Antibiotic Sensitivity Testing (AST), Modified Hodge Test (MHT) and Modified Carbapenem inactivation method (mCIM) were performed for detection of carbapenemase production and Congo red agar method (CRA) along with Microtitre plate method were performed for detecting biofilm production.
 Results: Among the clinical specimens cultured, growth positivity was 62.71%. E. coli was most predominant organism followed by K. pneumoniae (17.89%). Among the 110 K. pneumoniae, 57 were found to be carbapenemase producer. Majority of the carbapenemase producing K. pneumoniae were isolated from sputum (45.61%), in the specimen collected from age group 61-70 (28.07%) and in out-patient department (50.88%). Similarly, 65.45% K. pneumoniae out of 110 were found to be biofilm producer by Congo red agar method while among those 72, 73.59% isolates were found to be quantitatively biofilm producer in Microtitre plate assay. Out of 57 carbapenemase producer, 35.08% were strongly biofilm producer while among 53 carbapenemase non-producer 30.18% were strongly biofilm producer from Congo red agar method. Moreover, Microtitre plate assay evidenced that, out of 57 carbapenemase producer, 40.35% were highly biofilm producing and among the 15 carbapenemase nonproducer 66.66% were highly biofilm producer. 
 Conclusion: Biofilm formation is highly prevalent with varying degree of resistance among different antibiotics including carbapenems that further augments antibiotic resistance. The study showed carbapenemase producers are stronger biofilm producer than the non-carbapenemase producer. Therefore, it is recommended to identify biofilm formation among carbapenemase producers for effective choice of antibiotics.
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Shrestha, Bijay Kumar, and Jenish Shakya. "In-Vitro Evaluation of Biofilm and Hemolysis activity of Candida albicans Isolated from Oral Cavity." International Journal of Applied Sciences and Biotechnology 8, no. 4 (2020): 394–99. http://dx.doi.org/10.3126/ijasbt.v8i4.32971.
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Candida albicans is a member of the healthy human microflora, colonizing several niches in the body and can cause opportunistic infection under host debilitated and immunocompromised condition. The present study aimed to investigate the in-vitro hemolytic activity of C. albicans isolated from oral cavity and screen biofilm through three different methods. During the study, 200 oral rinse samples from general human population were analyzed in microbiology laboratory of Central Campus of Technology, Tribhuvan University, Hattisar, Dharan. Nepal. Candida albicans were isolated and identified by conventional microbiological procedures. The hemolytic activity was evaluated through two different Sabouraud dextrose broth media (SDB) containing 7% defibrinated human blood, one supplemented with 3% glucose (SDBwG) and the other without glucose (SDBwoG). The biofilm formation was screened through congo red agar, tube method and tissue culture plate method. In this present study, 42 (21%) isolates of Candida albicans were isolated from 200 oral rinse samples. Isolated Candida albicans exhibited mean hemolysis activity of 28.66% on human blood SDB without glucose and 43.55% on human blood SDB with 3% glucose. Tissue culture plate method was considered sensitive, specific and accurate method for quantitative screening of biofilm in comparison to tube method and congo red agar method. This research concluded that Candida albicans exhibited greater hemolytic activity in human blood with glucose (SDBwG) than in human blood without glucose (SDBwoG). This finding explains that an increased blood glucose concentration may contribute to increased hemolysis activity of Candida albicans that could play pathogenic role for inducing infection like oral candidiasis in debilitated host like diabetic patients. Tissue culture plate method can accurately screen biofilms than tube and congo red agar method.
 Int. J. Appl. Sci. Biotechnol. Vol 8(4): 394-399
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A. ABDULLAH, Zheen, and Aisha K. Ahmed. "SCREENING OF ANTIBIOTIC RESISTANCE BETWEEN BIOFILM PRODUCER AND NON-BIOFILM PRODUCER OF KLEBSIELLA PNEUMONIAE ISOLATES." MINAR International Journal of Applied Sciences and Technology 4, no. 4 (2022): 125–32. http://dx.doi.org/10.47832/2717-8234.13.11.
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Background: Klebsiella pneumoniae is a common cause of nosocomial infections. Antibiotic resistance and the ability to form biofilm, as two key virulence factors of K. pneumoniae, are involved in the persistence of infections. Aim: determine the rate of (10) antibiotics activity against isolated K. pneumoniae then evaluate the ability of isolated bacteria to produce biofilm by using Congo red agar method and studying the antibiotic resistance pattern between biofilm producer and non-biofilm producer of K. pneumoniae isolates. Material and methods: over a period of 4 months a total of 37 k. pneumoniae isolates were collected. Antibiotic susceptibility was determined by the disc diffusion method according to CLSI. Biofilm production was assessed by Congo red agar method. Results: Antibiotic susceptibility profile showed a variable level of resistance. The highest resistance was reported with tetracycline (56.8%). And the highest susceptibility was reported with imipenem (85.6%) and meropenem (85.6%). Biofilm by Congo red agar method was detected in 19 isolates with a percentage of (51.35%) however, 18 (48.65%) of isolates were non biofilm producer. The association of antibiotic resistance between biofilm-producing and non-biofilm-producing K. pneumoniae isolates was evaluated by the Chi-square test and the p-value was found to be 0.97 (statistically non-significant). Conclusion: our study revealed that the K. pneumoniae isolates differed in their responses against used antibiotics, moreover they are varied in their ability to produce biofilm and the resistance pattern between biofilm producer and non-producer was no significant. More studies are needed to characterize the pattern of antibiotic resistance between biofilm producer and non-biofilm producer of K. pneumoniae isolates and universal efforts should be increased to prevent the prevalence of multi- antibiotic resistant bacteria and eliminate the hospital born bacteria that are causing a great rise in mortality. Keywords: Klebsiella Pneumoniae, Antibiotic Resistance, Biofilm
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Öcal, Duygu, Alper Tekeli, and İştar Dolapçı. "Evaluation of Biofilm Formation in Coagulase Negative Staphylococci on Various Congo Red Agar Medium." Journal of Ankara University Faculty of Medicine 75, no. 1 (2022): 8–15. http://dx.doi.org/10.4274/atfm.galenos.2021.54366.
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Kaiser, Thaís Dias Lemos, Eliezer Menezes Pereira, Kátia Regina Netto dos Santos, Ethel Leonor Noia Maciel, Ricardo Pinto Schuenck, and Ana Paula Ferreira Nunes. "Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis." Diagnostic Microbiology and Infectious Disease 75, no. 3 (2013): 235–39. http://dx.doi.org/10.1016/j.diagmicrobio.2012.11.014.
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Blanco, Anna Rita, Andrea Sudano-Roccaro, Giovanna Carmela Spoto, Antonia Nostro, and Dario Rusciano. "Epigallocatechin Gallate Inhibits Biofilm Formation by Ocular Staphylococcal Isolates." Antimicrobial Agents and Chemotherapy 49, no. 10 (2005): 4339–43. http://dx.doi.org/10.1128/aac.49.10.4339-4343.2005.
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ABSTRACT Epigallocatechin gallate (EGCg), the main polyphenol component of green tea, has several antibacterial properties. Here we show that sub-MICs of EGCg appear to decrease slime production, therefore inhibiting biofilm formation by ocular staphylococcal isolates previously characterized for the presence of ica genes by the Congo red agar plate assay and for adhesion to microtiter plates.
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Florencio, Camila, Sonia Couri, and Cristiane Sanchez Farinas. "Correlation between Agar Plate Screening and Solid-State Fermentation for the Prediction of Cellulase Production by Trichoderma Strains." Enzyme Research 2012 (November 28, 2012): 1–7. http://dx.doi.org/10.1155/2012/793708.
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The viability of converting biomass into biofuels and chemicals still requires further development towards the reduction of the enzyme production costs. Thus, there is a growing demand for the development of efficient procedures for selection of cellulase-producing microorganisms. This work correlates qualitative screening using agar plate assays with quantitative measurements of cellulase production during cultivation under solid-state fermentation (SSF). The initial screening step consisted of observation of the growth of 78 preselected strains of the genus Trichoderma on plates, using microcrystalline cellulose as carbon source. The 49 strains that were able to grow on this substrate were then subjected to a second screening step using the Congo red test. From this test it was possible to select 10 strains that presented the highest enzymatic indices (EI), with values ranging from 1.51 to 1.90. SSF cultivations using sugarcane bagasse and wheat bran as substrates were performed using selected strains. The CG 104NH strain presented the highest EGase activity (25.93 UI·g−1). The EI results obtained in the screening procedure using plates were compared with cellulase production under SSF. A correlation coefficient (R2) of 0.977 was obtained between the Congo red test and SSF, demonstrating that the two methodologies were in good agreement.
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Pervin, Safirun, Bushra Jannat, and Sohana Al Sanjee. "Characterization of Rhizobia from Root Nodule and Rhizosphere of Lablab purpureus and Vigna sinensis in Bangladesh." Turkish Journal of Agriculture - Food Science and Technology 5, no. 1 (2017): 14. http://dx.doi.org/10.24925/turjaf.v5i1.14-17.743.
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Nitrogen fixation resulting from mutual symbiosis of rhizobia and cultivated legume plants is therefore critical to food security as it directly affects agricultural production. Biological Nitrogen Fixation (BNF) can be an important factor in sustainable agriculture.The isolation and identification of different slow growing and fast growing rhizobial strains from the nodules of two leguminous plant species. Symbiotic nitrogen fixing Rhizobium spp. was isolated from (Lablab purpureus and Vigna sinensis). Nodules samples were collected from plants growing in different Districts of Bangladesh and the Glucose-Peptone Agar (GPA), Congo red, Yeast Mannitol Agar (YMA) containing 2% NaCl were employed to make presumptive decisions on the recognition and classification of the isolated bacterial strains. All the isolates were found with poor absorption of dye Congo red and little or no growth on the media of GPA and without altering the pH. Almost all of the isolates exhibit growth on 2% NaCl, poor growth on GPA, thus confirming the rhizobia. After biochemical tests like catalase test and citrate utilization test isolates were confirmed as Rhizobia. The presence of rhizobia on root nodules of leguminous Plant. Not only the leguminous Plant but also the rhizosphere contains rhizobia which help in soil fertilization.
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Prakash, Prabhu, Richa Agarwal, Ekta Gupta, Ravinder Singh Rathore, Vishakha Ashopa, and Eshank Gupta. "To study drug resistance & biofilm production in gram negative isolates from clinical samples." Indian Journal of Microbiology Research 9, no. 3 (2022): 200–206. http://dx.doi.org/10.18231/j.ijmr.2022.036.
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Biofilm are groups of microorganism encased in a matrix of extracellular polysaccharide (slime), called polysaccharide intercellular adhesion (PIA). Bacteria commonly involved include , , , and . The present study was performed to identify antibiotic resistance pattern & their ability to form biofilm in gram negative clinical isolates. All clinical samples received in laboratory for microbial culture during study period of 12 months (2017 to 2018) were included in this study Antibiotic susceptibility testing, ESBL & MBL detection was done for clinical isolates. Biofilm productions were determined by Congo red agar method, Christenson’s Test Tube method and Tissue culture plate method. 327 gram negative isolates were detected. Maximum were (32.72%) followed by (28.44%), (16.51%), (16.51%), Citrobacter species (3.97%). Maximum isolates showed resistance to ampicillin (93.27%) followed by amoxiclave (87.46%), ceftazidime (74%). Out of 327 GNB isolates, biofilm produced by 64 (19.57%) isolates by Tissue culture plate (TCP) method, 38(11.62%) by Congo red agar (CRA) methods and 23 (7.03%) by Tube methods. Maximum biofilm were detected in (24.29%). There is increase prevalence of multidrug resistant& biofilm forming bacteria. The routine monitoring of multidrug resistance pattern & biofilm detection can be recommended in clinical laboratories to guide proper antibiotic treatment.
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Triveni, Alasthimannahalli Gangadhara, Mendem Suresh Kumar, Chavadi Manjunath, Channappa T. Shivannavar, and Subhaschandra M. Gaddad. "Biofilm formation by clinically isolated Staphylococcus Aureus from India." Journal of Infection in Developing Countries 12, no. 12 (2018): 1062–66. http://dx.doi.org/10.3855/jidc.10671.
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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.
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Ugwoke, MI, DA Machido, and MB Tijjani. "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." Journal of Applied Sciences and Environmental Management 23, no. 10 (2019): 1783–86. http://dx.doi.org/10.4314/jasem.v23i10.3.
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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
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Ezeagu, G. G., M. Fadayomi, and U. S. Rikiji. "Cellulolytic potentials of aspergillus oryzae and streptomyces griseus isolated from waste dump soil in Nile University Of Nigeria, Abuja." Science World Journal 19, no. 1 (2024): 189–93. http://dx.doi.org/10.4314/swj.v19i1.25.
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The potential of using microorganisms as biological sources of industrially economic enzymes has stimulated interest in the exploitation of extracellular enzymatic activity in several microorganisms. The aim of this research is to assess the cellulose degrading potentials of two microorganisms, Aspergillus oryzae and Streptomyces griseus using cellulose Congo red agar media. Soil sample collected from waste dump was serially diluted and inoculated in starch casein agar and SDA to isolate S. griseus and A. oryzae respectively. To assess their potentials to utilize cellulose, each of the two microorganisms was inoculated on cellulose Congo-red media and incubated at 30 ºC for 7days. A zone of clearing around the colonies after incubation confirms the secretion of extracellular cellulase, and was used as an indication for cellulose utilization. The zone of clearing was measured with a meter rule. In the results obtained, both microorganisms demonstrated cellulose utilization ability with Aspergillus oryzae showing a zone of clearing of 30.50 ± 0.50 mm while Streptomyces griseus showed a wider zone of clearing of 60.00 ± 1.00 mm. The results indicate that both microorganisms can be potent producers of the enzyme cellulase, with Streptomyces griseus having a higher cellulase-producing ability.
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S. El. Naghy, Wageih, Sarah A. Hamam, Tamer A. Wasfy, and Sara M. Samy. "Detection of Biofilm Formation by Different Bacterial Isolates of Contact Lens." Egyptian Journal of Medical Microbiology EJMM29, no. 4 (2020): 93–100. http://dx.doi.org/10.51429/ejmm29412.
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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.
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Garba, Samira. "Detection of biofilm formation in multi-drug resistant Staphylococcus aureus among clinical isolates in Zaria metropolis, Kaduna, Nigeria." Journal of Current Biomedical Research 3, no. 2, March-April (2023): 878–88. http://dx.doi.org/10.54117/jcbr.v3i2.5.
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Antimicrobial resistance are associated with several virulence factors such as biofilm. The biofilm formation ability of S. aureus complicates effective therapeutic management of infections. In this research, we intend to observe biofilm formation in S. aureus using two in vitro phenotypic methods: tissue culture plate and Congo red agar as well as biofilm associated genes Ica A B & D and 16SrRNA. A total of 150 presumptive coagulase positive staphylococcal isolates from all specimens (blood, urine, high vaginal swab, wound swab, ear swab, endocervical swab) submitted to the microbiology laboratory of the selected hospitals were collected. Seventy-three (73) or 48.7% of the isolates were identified as coagulase positive Staphylococcus using microbiology standard methods (tube coagulase test, growth on mannitol salt agar & Dnase agar). Using microgen staph identification kit, the isolates were further characterized into various Staph spp. Disc agar diffusion method was used for eleven (11) antibiotics susceptibility test while vancomycin screening agar and BMD (Broth Micro dilution) MIC test was used for vancomycin susceptibility evaluation. Indirect observation of biofilm was performed using congo red agar (CRA) and Microtitre plate assay method. PCR and sequencing were conducted on the isolates to molecularly detect the presence of 16SrRNA, IcaA, IcaB, IcaD genes. Susceptibility of S aureus (21%) isolates tested against 12 different categories of antibiotics shows 4(27%) not multidrug resistant (MDR), 7(47%) were MDR and 4 (27%) were extensively drug resistant (XDR). High percentage (74%) of Multiple antibiotic resistant index at ≥0.3 suggested that the isolates originated from an environment where antibiotics are often used. Biofilm assessment of MDR S aureus isolates revealed that 9(82%) and 11 (100%) of the isolates were biofilm formers using Congo Red Agar method and Microtiter plate assay respectively. All isolates amplified with 16SrRNA primer signifying all are S. aureus, 100% of the isolates amplified with IcaA, 90% isolates amplified with IcaD, and 50% isolates amplified with IcaB. We can conclude that there is high prevalence of Biofilm forming S. aureus strains in the hospitals studied. This result should be born in mind by clinicians while prescribing medication for S. aureus positive conditions. Hospital managements should be proactive in preventing the preservation of the biofilm formers in their hospital environment through adequate sanitary and disinfection protocols. It is also necessary for clinician to note that nitrofurantoin, linezolid and vancomycin were most effective against the biofilm producing S. aureus and microtitre plate assay method was observed to be most reliable assay in the study of biofilm.
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Priyanka, Paul Biswas, Akhter Kahkashan, Fidai Farhaan, Singh Anamika, Sen Aninda, and Dey Akoijam Sangeeta. "Detection of Biofilm Formation and Virulence Markers Amongst the Cons Isolates in a Tertiary Care Center in Bihar." International Journal of Pharmaceutical and Clinical Research 13, no. 6 (2021): 269–77. https://doi.org/10.5281/zenodo.14215252.
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<strong>Background: </strong>The aim of the study was to speciate Coagulase negative staphylococci [CoNS] from blood samples, ascertain the antibiogram and to determine the expressions of these virulence markers and biofilm production in the CONS isolates. <strong>Materials and Methods: </strong>A total of 350 blood samples were collected from clinically suspected cases of BSI [during the fever spikes] for routine blood culture. A panel of standard biochemical tests was used to identify the CoNS upto species level. Phenotypic detection of virulence markers was done by standard test. Biofilm production was screened by tissue culture plate [TCP], Tube method [TM] & Congo red agar [CRA] and brain heart infusion agar [BHIA] with 6% sucrose method. Antibiogram was detected by modified Kirby Bauer method as per CLLSI guidelines. <strong>Results:</strong> 32.3% [45/139] isolates produced biofilm by standard tissue culture plate [TCP] assay. 5.7% [8/139] strains by tube method [TM] method followed by 2.1% [3/139] each by and Congo red agar [CRA] method & brain heart infusion agar [BHI] 6% suc method were strong biofilm producers in comparison to TCP method 15.1% [21/139]. The sensitivity & specificity of TA method was [53.3% & 81.9%]. However, sensitivity of CRA & BHIsuc6% method were much lower being, 28.8% & 20.0% whereas the specificity was 93.6% & 89.3%.. Production of DNase, lipase, caesinase and gelatinase was common to all the seven species, DNase being more common in of Staphylococcus epidermidis 42.8% [15/35]. <strong>Conclusions: </strong>The TCP method was found to be most sensitive, accurate and reproducible screening method for detection of biofilm formation. Biofilm forming capacity and elaboration of various virulence determinants followed by multi-drug resistance by CONS will facilitate its colonizing ability. Hence importance should be laid for routine identification of CONS and determining its antimicrobial susceptibility pattern.
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Jebril, Nadia Mahmoud Tawfiq. "Evaluation of two fixation techniques for direct observation of biofilm formation of Bacillus subtilis in situ, on Congo red agar, using scanning electron microscopy." June-2020 13, no. 6 (2020): 1133–37. http://dx.doi.org/10.14202/vetworld.2020.1133-1137.
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Background and Aim: Direct observation, scanning electron microscopy (SEM) is a common method used for the observations of biofilms. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide) (EDC) fixation method has proven to be a valuable fixation method in the observation of these biofilms. Still, it entails a method of biofilm fixation that can damage slim structures, leading to the impossible observation of biofilm development. In contrast, alcian blue and lysine (ABL) fixation technique appears more glycocalyx of biofilm, fully preserved samples, which may provide much insight into the development of B. subtilis biofilms. Materials and Methods: Here, the evaluation of the fixation of ABL technique for the study of B. subtilis biofilms was carried out in situ, on Congo red agar. In doing so, the comparison to commonly use conventional EDC technique for sample fixation, and observation was carried out. Observations were based on SEM over 30 samples. Results: Overall, ABL technique provided excellent observation of biofilms formed in situ, on Congo red agar, and revealed slime structures, which have not been observed, much in standard EDC fixation or earlier in other studies of these biofilms in B. subtilis. Conclusion: This study reported the appropriate use of ABL in the fixation technique for the preservation of biofilm of B. subtilis.
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Parrish, N. M., C. G. Ko, J. D. Dick, P. B. Jones, and J. L. E. Ellingson. "Growth, Congo Red Agar Colony Morphotypes and Antibiotic Susceptibility Testing of Mycobacterium avium subspecies paratuberculosis." Clinical Medicine & Research 2, no. 2 (2004): 107–14. http://dx.doi.org/10.3121/cmr.2.2.107.
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M. Raoof, Waad, Amera M. Mohamad, and Aisha W. Al-Omari. "Detection of Biofilm Formation in some Pathogenic Bacteria Using Tube and Congo Red Agar Methods." Rafidain Journal of Science 24, no. 12 (2013): 55–65. http://dx.doi.org/10.33899/rjs.2013.80268.
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Saxena, Naveen, Deepak Maheshwari, Divya Dadhich, and Savita Singh. "EVALUATION OF CONGO RED AGAR FOR DETECTION OF BIOFILM PRODUCTION BY VARIOUS CLINICAL CANDIDA ISOLATES." Journal of Evolution of Medical and Dental Sciences 3, no. 59 (2014): 13234–38. http://dx.doi.org/10.14260/jemds/2014/3761.
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Alamri, Aisha M., Afnan A. Alsultan, Mohammad A. Ansari, and Amani M. Alnimr. "Biofilm-Formation in Clonally Unrelated Multidrug-Resistant Acinetobacter baumannii Isolates." Pathogens 9, no. 8 (2020): 630. http://dx.doi.org/10.3390/pathogens9080630.
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This study analyzed the genotype, antibiotic resistance, and biofilm formation of Acinetobacter baumannii strains and assessed the correlation between biofilm formation, antibiotic resistance, and biofilm-related risk factors. A total of 207 non-replicate multi-drug-resistant A. baumannii strains were prospectively isolated. Phenotypic identification and antimicrobial susceptibility testing were carried out. Isolate biofilm formation ability was evaluated using the tissue culture plate (TCP), Congo red agar, and tube methods. Clonal relatedness between the strains was assessed by enterobacterial repetitive intergenic consensus-PCR genotyping. Of the 207 isolates, 52.5% originated from an intensive care unit setting, and pan resistance was observed against ceftazidime and cefepime, with elevated resistance (99–94%) to piperacillin/tazobactam, imipenem, levofloxacin, and ciprofloxacin. alongside high susceptibility to tigecycline (97.8%). The Tissue culture plate, Tube method, and Congo red agar methods revealed that 53.6%, 20.8%, and 2.7% of the strains were strong biofilm producers, respectively, while a significant correlation was observed between biofilm formation and device-originating respiratory isolates (p = 0.0009) and between biofilm formation in colonized vs. true infection isolates (p = 0.0001). No correlation was detected between antibiotic resistance and biofilm formation capacity, and the majority of isolates were clonally unrelated. These findings highlight the urgent need for implementing strict infection control measures in clinical settings.
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Almeida, Juliana Afonso de, Caroline Espíndola de Barros, Gustavo Henrique da Silva Ayres, et al. "Resistance profile from staphylococcus aureus and pseudomonas aeruginosa obtained from tracheostomized children in the four seasons of the year." Brazilian Journal of Development 8, no. 12 (2022): 81108–33. http://dx.doi.org/10.34117/bjdv8n12-285.
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Objective: The objective of this study was to microbiologically characterize the tracheal secretion of tracheostomized children, evaluate biofilm formation and study the phenotypic and molecular profile of the antimicrobial resistance of Sthapylococcus aureus and Pseudomonas aeruginosa isolates. Methods: 88 samples of tracheal secretions were collected. The material was processed to perform phenotypic tests and bacterial identification. Tests were used to identify biofilms using Congo red agar test and microdilution in a 96-well plate, and the qPCR method was used to verify resistance. Results: Twelve Staphylococcus aureus samples and 30 Pseudomonas aeruginosa were isolated of pediactric tracheostomized patients. All the S. aureus samples were positive to biofilm formation in Congo red agar test. In the antibiogram test, S. aureus showed resistance to seven antimicrobials. Regarding the identification of resistance genes, blaZ was amplified in 57.1% and mecA in 28.6% of the isolated S. aureus. Pseudomonas aeruginosa showed blaOXA with 66,7% e blaKPC with 58,3%. In plasmid DNA, blaNDM stood out with 58,3% positive. Conclusions: The control of resistant bacteria involved in biofilms in the stoma of tracheostomized patients is a great challenge, since the simple cannula change does not always allow the control of the microbiota, which increases the vulnerability of patients to future respiratory complications.
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HART, R. A., O. MO, F. BORIUS, and D. Y. C. FUNG. "COMPARATIVE ANALYSIS OF TRYPAN BLUE AGAR AND CONGO RED AGAR FOR THE ENUMERATION OF YEAST AND MOLD USING THE HGMF SYSTEM." Journal of Food Safety 11, no. 3 (1990): 227–30. http://dx.doi.org/10.1111/j.1745-4565.1990.tb00054.x.
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Cangelosi, Gerard A., Julie S. Do, Robert Freeman, John G. Bennett, Makeda Semret, and Marcel A. Behr. "The Two-Component Regulatory System mtrAB Is Required for Morphotypic Multidrug Resistance in Mycobacterium avium." Antimicrobial Agents and Chemotherapy 50, no. 2 (2006): 461–68. http://dx.doi.org/10.1128/aac.50.2.461-468.2006.
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ABSTRACT Clinical isolates of the opportunistic pathogen Mycobacterium avium complex (MAC) undergo a reversible switch between red and white colony morphotypes on agar plates containing the lipoprotein stain Congo red. Compared to their isogenic red counterparts, white morphotypic variants are more virulent and more resistant to multiple antibiotics. This report shows that the two-component regulatory system mtrAB is required for the red-to-white switch as well as for other morphotypic switches of MAC. A mutant with a transposon insertion in the histidine protein kinase gene mtrB was isolated from a morphotypically white parent clone. The mutant resembled a naturally occurring red morphotypic variant in that it stained with Congo red, was sensitive to multiple antibiotics, and was permeable by a fluorescent DNA stain. However, it differed from a red variant in that it could not switch to the white or transparent morphotype, and it could not survive intracellularly within macrophage-like cells. Transcomplementation with a cloned wild-type mtrB gene restored to the mutant the ability to form impermeable, drug-resistant white and transparent variants. Quantitative reverse transcriptase PCR showed that mtrB was required for the normal expression of cell surface Mce proteins, some of which are up-regulated in the red-to-white switch. The results indicate that mtrAB functions in regulating the composition and permeability of mycobacterial cell walls and plays a role in the reversible colony type switches of MAC.
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Solaiman, ARM, GMA Hossain, and MAB Mia. "Effect of Rhizobium on Growth and Biomass Production of Rice." Bangladesh Journal of Microbiology 28, no. 2 (2012): 64–68. http://dx.doi.org/10.3329/bjm.v28i2.11818.
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To characterize twenty Rhizobium strains isolated from nodules of lentil, grasspea and chickpea, an experiment was conducted in the Soil Microbiology laboratory of the Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh. The isolates were tested for growth on Congo red Yeast Extract Mannitol (YEM) agar, peptone glucose agar, and YEM agar containing Bromothymol blue (BTB). All the strains except Ls 12 absorbed Congo red dye very weekly. Except Le 2 strains, isolated from lentil showed no growth in peptone glucose agar. All the strains isolated from grasspea and chickpea showed moderate growth on this medium. All the strains were fast-growing and showed acidic reaction on YEM agar medium. Among the strains isolated from lentil, Le 1, Le 2 and Le 4 produced moderate turbidity while Le 3, Le 6, Le 7 and Le 8 produced high turbidity in YEM broth. All the strains isolated from grasspea except Ls 3 and chickpea except Ca 1 produced moderate turbidity in YEM broth medium. Strains Ls 3 and Ca 1 produced high turbidity. To assess the effect of ten of these Rhizobium isolates viz. Le 1, Le 4, Le 6, Le 8, Ls 1, Ls 2, Ls 6, Ls 7, Ca 3 and Ca 4 on growth and biomass production of rice, a follow-up experiment was conducted in the same laboratory. Root length of rice was significantly increased over control (without inoculation) due to inoculation with different Rhizobium strains. The highest root length (9.63 cm) was obtained by inoculation with strain Ls 6 isolated from lentil. All the Rhizobium strains produced significantly higher shoot length, fresh and dry biomass over control. The highest shoot length (16.50 cm), fresh biomass (138.3 mg) and dry biomass (27.75 mg) were also obtained from the strain Ls 6. DOI: http://dx.doi.org/10.3329/bjm.v28i2.11818 Bangladesh J Microbiol, Volume 28, Number 2, December 2011, pp 64-68
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Osungunna, Michael O., and Grace O. Onawunmi. "Antibiotic resistance profiles of biofilm-forming bacteria associated with urine and urinary catheters in a tertiary hospital in Ile-Ife, Nigeria." Southern African Journal of Infectious Diseases 33, no. 3 (2018): 80–85. http://dx.doi.org/10.4102/sajid.v33i3.12.
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Background: Microorganisms that infect humans differ in pathogenesis, virulence factors and antimicrobial resistance profiles. In natural settings, bacterial cells are most often found in close association with surfaces and interfaces, in the form of multicellular aggregates commonly called biofilms. Given their ubiquity and importance in the microbial world, it is hardly surprising that biofilms have attracted the attention of the scientific community. Biofilm formation on medical implant devices such as catheters is also a major problem that is closely tied to the adhesion and resistance-related abilities of the biofilm.Methodology: The ability of 216 bacterial isolates from mid-stream urine (100), catheter-stream urine (52) and catheter tips (64) to form biofilms was investigated using the tissue culture plate method, the tube and Congo red agar methods as well as their antibiotic resistance profiles using the agar disc diffusion method.Results: These revealed that Klebsiella spp. was the predominant bacterial genera accounting for 45.8% of the total isolates. A total of 50 isolates were biofilm-formers with 22% identified by the tissue culture plate method and 78% identified by the Congo red agar method. Klebsiella spp. had the highest ability to form biofilm while antibiotic resistance profiles showed all the biofilmformers to be multiply antibiotic resistant with least resistance to ofloxacin.Conclusion: It can therefore be concluded that some bacterial isolates associated with urinary tract infections have a propensity to form biofilm, thereby becoming multiply antibiotic resistant, and ofloxacin remains the antibiotic of choice in the treatment of such infections.
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Medegar, Shobha, Pooja Mansabdar, and Asha B. Patil. "An evaluation of three different biofilm detection methods in orthopedic implant associated infections and its implication in healthcare system." IP International Journal of Medical Microbiology and Tropical Diseases 8, no. 1 (2022): 63–68. http://dx.doi.org/10.18231/j.ijmmtd.2022.014.
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: Biofilms are defined as microbially derived sessile communities characterized by the cells that are irreversibly attached to a substratum. These organisms have been associated with implant associated infectionsThe present study is therefore undertaken 1. To compare the three screening methods used for detection of biofilm formation (Tissue culture plate method, Tube method and Congo red agar method) and 2. To correlate the antibiotic resistance pattern of biofilm producers and non biofilm producers. : A prospective study was carried out in the department of microbiology, on all orthopedic implant associated infections from September 2015 to July 2016 on aspirated pus samples. A total of 120 non repetitive clinical isolates were taken and subjected to biofilm detection. All the bacterial isolates were identified by standard biochemical tests. Antibiotic susceptibility test of bacterial isolates was performed by Kirby Bauer disk diffusion method according to Clinical Laboratory Standard Institute (CLSI) guidelines on Muller Hinton Agar (MHA). A reference strain of Staphylococcus epidermidis ATCC 35984(positive biofilm producer) and Staphylococcus epidermidis ATCC 12228(non biofilm producer) were used as controls. Tissue culture plate method was considered as gold standard as it detected 25% biofilm producers which included 12% weak biofilm producers which was missed by congo red agar (CRA) and tube method (TM). Also, this correlated to methicillin resistant staphylococcus aureus (MRSA), ESBL, amp-c and MDR pattern of antibiogram. : TCP is considered as most reliable screening method for biofilm detection and should be routinely employed to prevent treatment failures as these organisms are intrinsically resistant to many antimicrobials leading to implant associated infections.
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Uhlich, Gaylen A., Peter H. Cooke, and Ethan B. Solomon. "Analyses of the Red-Dry-Rough Phenotype of an Escherichia coli O157:H7 Strain and Its Role in Biofilm Formation and Resistance to Antibacterial Agents." Applied and Environmental Microbiology 72, no. 4 (2006): 2564–72. http://dx.doi.org/10.1128/aem.72.4.2564-2572.2006.
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ABSTRACT In a previous study, we identified Congo red-binding and -nonbinding phase variants of Escherichia coli serotype O157:H7 strain ATCC 43895. The Congo red-binding variant, strain 43895OR, produced a dry, aggregative colony that was similar to the red, dry, and rough (rdar) phenotype characteristic of certain strains of Salmonella. In contrast, variant 43895OW produced a smooth and white colony morphology. In this study, we show that, similar to rdar strains of Salmonella enterica serovar Typhimurium, strain 43895OR forms large aggregates in broth cultures, firm pellicles at the air-medium interface on glass, and dense biofilms on glass and polystyrene. However, unlike S. enterica serovar Typhimurium, strain 43895OR does not stain positive for cellulose production. When strain 43895OR was fixed on agar, scanning electron microscopy showed cells expressing extracellular matrix (ECM) containing curli fibers. Strain 43895OW was devoid of any ECM or curli fibers on agar but showed expression of curli fibers during attachment to glass. Strain 43895OR produced >4-fold-larger amounts of biofilm than strain 43895OW on polystyrene, glass, stainless steel, and Teflon; formation was >3-fold higher in rich medium than in nutrient-limited medium. Biofilm-associated cells of both strains showed statistically greater resistance (P < 0.05) to hydrogen peroxide and quaternary ammonium sanitizer than their respective planktonic cells. This study shows that the rdar phenotype of E. coli O157:H7 strain 43895OR is important in multicellular growth, biofilm formation, and resistance to sanitizers. However, the lack of cellulose production by strain 43895OR indicates important differences in the ECM composition compared to that of Salmonella.