scholarly journals In-Vitro Evaluation of Biofilm and Hemolysis activity of Candida albicans Isolated from Oral Cavity

2020 ◽  
Vol 8 (4) ◽  
pp. 394-399
Author(s):  
Bijay Kumar Shrestha ◽  
Jenish Shakya
Keyword(s):  
Tissue Culture ◽  
Candida Albicans ◽  
Human Blood ◽  
Hemolytic Activity ◽  
Congo Red ◽  
Plate Method ◽  
Oral Rinse ◽  
Culture Plate ◽  
Tissue Culture Plate

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

scholarly journals Antibiotic resistance profiles of biofilm-forming bacteria associated with urine and urinary catheters in a tertiary hospital in Ile-Ife, Nigeria

2018 ◽  
Vol 33 (3) ◽  
pp. 80-85
Author(s):  
Michael O. Osungunna ◽  
Grace O. Onawunmi
Keyword(s):  
Tissue Culture ◽  
Antibiotic Resistance ◽  
Congo Red ◽  
Bacterial Isolates ◽  
Plate Method ◽  
Antibiotic Resistant ◽  
Culture Plate ◽  
Tissue Culture Plate ◽  
Tract Infections ◽  
Klebsiella Spp

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.

scholarly journals Detection of Biofilm Producing Staphylococci among Different Clinical Isolates and Its Relation to Methicillin Susceptibility

2018 ◽  
Vol 6 (8) ◽  
pp. 1335-1341 ◽  
Author(s):  
Rania M. Abdel Halim ◽  
Nevine N. Kassem ◽  
Basma S. Mahmoud
Keyword(s):  
Tissue Culture ◽  
Congo Red ◽  
Biofilm Formation ◽  
Methicillin Resistance ◽  
Screening Method ◽  
Culture Plate ◽  
Tissue Culture Plate ◽  
Blood Specimens ◽  
Phenotypic Methods

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.

scholarly journals Assessment of Biofilm Formation among the Clinical Isolates of Escherichia coli in a Tertiary Care Hospital

2020 ◽  
pp. 26-32
Author(s):  
Bedobroto Biswas ◽  
Naik Shalini Ashok ◽  
Deepesh Nagarajan ◽  
Md Zaffar Iqubal
Keyword(s):  
Escherichia Coli ◽  
Tissue Culture ◽  
Biofilm Formation ◽  
Tertiary Care ◽  
Clinical Samples ◽  
Care Hospital ◽  
Plate Method ◽  
Culture Plate ◽  
Tissue Culture Plate

Aims: Identification and grading of the Escherichia coli according to their biofilm production capability. Study Design:  Cross-sectional study. Place and Duration of Study: This was conducted in Department Microbiology at M.S. Ramaiah Medical college and Hospital, Bengaluru from March 2017 to August 2017. Methodology: A total of 55 non repetitive Escherichia coli isolates were identified from various clinical samples like urine, pus ,tissue and peritoneal fluids .All the organisms were isolated in pure culture and biofilm formation was detected in vitro by Gold standard TCP (Tissue culture plate) method. Organisms were incubated for an extended period of 48 hours and the biofilms were detected by acetone alcohol elution method. Organisms were categorized as strong, moderate, weak and no biofilm producers based on the obtained OD value of the elute. Results: Majority of the isolates of Escherichia coli were obtained from catheterized urine culture (67.03%) followed by pus (25.50%).Most of the isolates were capable of forming biofilm in vitro by tissue culture plate method except a few (9.1%). 40% of the isolates were strong biofilm formers which had >4 ODC. 25.5% showed medium biofilm-forming capability and rest 25.5% showed weak biofilm formations in vitro. Conclusion: The ability to form biofilm from a species can give us a better understanding of the biofilm-related infections pertaining to the particular group. Detection of biofilms remains a most important determinant to approximate the incidence of such infections. Categorization of organisms according to their biofilm formation may help us understand the frequency of biofilm-associated infections, and thus take necessary precautions to avoid the problem. Further studies involving the detection of biofilm may be conducted and the tests can be implemented in routine diagnostic microbiology to assess the usefulness of the methods in detection of biofilm-related infections.

scholarly journals ANTIPLAQUE EFFICACY OF GANODERMA LUCIDUM TOOTHPASTE - AN IN VITRO STUDY

2018 ◽  
Vol 11 (11) ◽  
pp. 300
Author(s):  
Prabhjeet S ◽  
Meena A K ◽  
Jesil M
Keyword(s):  
Tissue Culture ◽  
Ganoderma Lucidum ◽  
In Vitro Study ◽  
Vitro Study ◽  
Distilled Water ◽  
Culture Plate ◽  
Significant Difference ◽  
Tissue Culture Plate

Objective: The objective of the study was to evaluate the efficacy of Ganoderma lucidum toothpaste as an antiplaque agent and to compare its efficacy with herbal toothpaste and mouthwash.Methods: Pooled saliva was collected in a sterile container from the volunteers after taking the consent. Tissue culture plate with 12 (3 × 4) wells was chosen. Pooled saliva of 20 mL was added to each well using the micropipette and was kept in the incubator at 37°C for 72 h. After 72 h, saliva was removed without touching the walls or the base of the wells. Each row was treated either with slurry prepared with Ganoderma/herbal/Colgate total toothpaste or herbal/chlorhexidine mouthwash/distilled water. One row of wells was kept as a control using erythrosine dye. After 30 s, all the wells were rinsed with distilled water. Erythrosine dye was added to all the wells, kept for 30 s, and rinsed with distilled water. The tissue culture plate was kept in the ELx800MS machine (ELISA reader) which was set at 540 nm, and the readings were obtained.Results: The results showed that G. lucidum toothpaste slurry reduced plaque than herbal and chlorhexidine mouthwash. However, there was no significant difference in plaque reduction between herbal and G. lucidum toothpaste slurries.Conclusion: The present study concluded that G. lucidum had better antiplaque efficacy than herbal toothpaste, herbal mouthwash, and chlorhexidine mouthwash.

scholarly journals Effect of Glucose Induction on Biofilm Density in Clinical Isolate Acinetobacter baumannii Patients in Intensive Care Unit of Dr. Soetomo Hospital, Surabaya

2020 ◽  
Vol 56 (2) ◽  
pp. 118
Author(s):  
Wira W Lindarto ◽  
Eddy Bagus Wasito ◽  
Kartuti Debora
Keyword(s):  
Tissue Culture ◽  
Acinetobacter Baumannii ◽  
Clinical Isolate ◽  
Growth Media ◽  
Plate Method ◽  
Culture Plate ◽  
Tissue Culture Plate ◽  
Glucose Induction ◽  
Sugar Levels ◽  
Effect Of Glucose

This study aimed to analyze the effect of glucose induction on the clinical isolate biofilm density of Acinetobacter baumannii. Thirteen clinical isolates of A. baumannii non biofilm forming were collected from non-DM patients who were treated at the ICU of Dr. Soetomo Hospital, Surabaya, was treated with the addition of 0.08% glucose, 0.15% glucose, 0.2% glucose, and 0.4% glucose in TSB growth media, followed by biofilm density examination with Tissue Culture Plate Method (TCPM) using 96 wells flatbottomed polyesterene tissue culture plate and read by autoreader ELISA with a wavelength of 630 nm (OD630). Biofilm density obtained was analyzed using ANOVA statistical analysis. The results of OD630 showed that the biofilm density increased significantly at the addition of 0.2% and 0.4% glucose. There was a significant increase in biofilm density at the addition of 0.2% and 0.4% glucose so that the management of blood sugar levels in ICU patients was needed before and when medical devices were installed.

scholarly journals Evaluasi Metode Skrining Biofilm Enterococcus faecalis Isolat Klinik Kateter Urin

2019 ◽  
Author(s):  
Wani Devita Gunardi ◽  
Mohamad Yanuar Prasetyo Nugroho ◽  
Elisabeth D. Harahap
Keyword(s):  
Tissue Culture ◽  
Enterococcus Faecalis ◽  
Congo Red ◽  
National Healthcare ◽  
National Healthcare Safety Network ◽  
Culture Plate ◽  
Tissue Culture Plate

Laporan National Healthcare Safety Network dari tahun 2006 - 2008, menunjukan penyebab paling umum kedua infeksi saluran kemih (ISK) terkait kateter adalah genus Enterococcus setelah Eschericia coli. Pada infeksi saluran kemih terkait kateter urin, faktor yang berperan penting dalam patogenesis infeksi ini, yaitu: pembentukan biofilm pada kateter urin. Peranan Enterococcus faecalis sebagai penyebab infeksi saluran kemih berkaitan dengan kemampuannya dalam membentuk biofilm. Ada beberapa variasi metode untuk mendeteksi pembentukan biofilm seperti Tissue Culture Plate (TCP), Tube method (TM), dan Congo Red Agar (CRA). Tujuan penelitian ini untuk mendapatkan metode deteksi pembentukan biofilm dari E. faecalis yang tepat, cepat, dan mudah dilakukan. Total tigabelas isolat bakteri E. faecalis yang didapat dari hasil isolasi kultur kateter urin dilakukan uji deteksi pembentukan biofilm dengan metode TCP sebagai baku emas dan CRA sebagai pemeriksaan pembanding. Hasilnya, didapatkan 61,5% dan 69,2% bakteri E. faecalis mampu menghasilkan biofilm menggunakan metode TCP dan CRA. Hasil uji diagnostik metode CRA dibandingkan dengan metode TCP untuk deteksi pembentukan biofilm, didapatkan sensitivitas dan spesifisitas dari CRA sebesar 75% dan 40%. CRA merupakan metode yang cepat dan mudah untuk dilakukan, namun memiliki spesifitas yang kurang baik. Hal ini dapat disebabkan pembacaan hasil yang bersifat subjektif dan menimbulkan kesalahan paralaks. Kata kunci : Enterococcus faecalis, Kateter Urin, Biofilm.

scholarly journals Biofilm-Formation in Clonally Unrelated Multidrug-Resistant Acinetobacter baumannii Isolates

Pathogens ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 630 ◽  
Author(s):  
Aisha M. Alamri ◽  
Afnan A. Alsultan ◽  
Mohammad A. Ansari ◽  
Amani M. Alnimr
Keyword(s):  
Tissue Culture ◽  
Antibiotic Resistance ◽  
Acinetobacter Baumannii ◽  
Congo Red ◽  
Biofilm Formation ◽  
Multidrug Resistant ◽  
Control Measures ◽  
Infection Control Measures ◽  
Culture Plate ◽  
Tissue Culture Plate

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.

scholarly journals Phenotypic and genotypic characterization of biofilm producing clinical coagulase negative staphylococci from Nepal and their antibiotic susceptibility pattern

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Sarita Manandhar ◽  
Anjana Singh ◽  
Ajit Varma ◽  
Shanti Pandey ◽  
Neeraj Shrivastava
Keyword(s):  
Tissue Culture ◽  
Congo Red ◽  
Antibiotic Susceptibility ◽  
Clinical Samples ◽  
Susceptibility Pattern ◽  
Coagulase Negative Staphylococci ◽  
Antibiotic Susceptibility Pattern ◽  
Biofilm Production ◽  
Culture Plate ◽  
Tissue Culture Plate

Abstract Background Coagulase-negative staphylococci (CNS) survive as commensals of skin, anterior nares and external canals of human and were regarded as non-infectious pathogens. However, they are emerging as a major cause of nosocomial infectious due to their ability to form biofilms and high resistance to several classes of antibiotics. This study examines the biofilm forming abilities of 214 clinical CNS isolates using phenotypic and genotypic methods, and determines their antibiotic susceptibility patterns. Methods A total of 214 clinical isolates collected from different clinical samples were identified as CNS and their antibiotic susceptibility determined by CLSI guidelines. The biofilm forming ability of all isolates was determined by three phenotypic methods; Congo red agar (CRA) method, tube adherence method (TM) and tissue culture plate (TCP) method and by genotypic method for the detection of icaAD genes. Results Among all the isolates, S. epidermidis (57.5%) was found the most frequently, followed by S. saprophyticus (18.7%), S. haemolyticus (11.2%), S. hominis (7%), and S. capitis (5.6%). Antibiotic susceptibility pattern demonstrated 91.6% isolates were resistant to penicillin and 66.8% to cefoxitin while 91.1% isolates were susceptible to chloramphenicol. Constitutive and inducible clindamycin resistant phenotype as measured by D-test was seen among 28% and 14.5% of isolates respectively. Tissue culture plate method detected biofilm production in 42.1% isolate followed by 31.8% through tube method while 20.1% isolates were found to produce slime in Congo red agar method. The genotypic assay revealed presence of icaA and icaD genes in 19.2% isolates. Conclusion The study shows a high prevalence of biofilm formation and inducible clindamycin resistance in CNS isolates, indicating the importance of in-vitro biofilm production test and D-test in routine laboratory diagnostics. Implementation of efficient diagnostic techniques for detection of biofilm production in clinical samples can help manage staphylococcal infections and minimize risks of treatment failures in hospitals.

scholarly journals Comparative study of antibiofilm activity of lime juice and lithium dioxide nanoparticles against Salmonella isolated from local cheese

2018 ◽  
Vol 12 (2) ◽  
pp. 82-94
Author(s):  
Zainab Zamel Khalaf
Keyword(s):  
Tissue Culture ◽  
Synergistic Effect ◽  
Comparative Study ◽  
Congo Red ◽  
Microbial Contamination ◽  
Antibiofilm Activity ◽  
Plate Method ◽  
Lime Juice ◽  
White Cheese ◽  
Tissue Culture Plate

      Thirty specimens of fresh white cheeses, presented for sale in different markets at different cities of Iraq were analyzed microbiologically in this study.   Isolates of Salmonella, included in the specimens collected from November 2017 to January 2018, were investigated. Capacity for biofilm producing was demonstrated by two method, Tissue culture plate method (TCP) and Congo red agar (CRA).After that the antibiofilm activity of lime extract and LiO2NPs was studied as each one of them alone and then the synergistic effect was done by TCP method. The results showed that all Salmonella isolates produce biofilm but in different degrees. The results also displayed that Lime extract and LiO2NPs had antibiofilm effect against Salmonella when used alone and when the combination done between each one of these materials. In conclusion, it was observed that the specimens of fresh white cheese included in this study contained microbial contamination at a health-threatening level but we can eliminate this contamination by using lime extract and LiO2NPs.

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