Characterization of Biofilm production in XDR Proteus mirabilis isolated from Baghdad city hospitals

Objective(s): P. mirabilis is considered as extensive drug resistant pathogen in many studies as it can resist complex antibiotic regimes, such pathogen can be threat to public health, especially when it has the ability to produce biofilm. Therefore, biofilm production was characterized among XDR P. mirabilis local isolates in this research. Material and Methods: 100 P. mirabilis isolated from wound infections from patients admitted to Baghdad hospitals. They were identified using biochemical test and Vitek2 system. The MIC test for antibiotic sensitivity was done by Vitek 2 automated system. Biofilm production was identified phenotypically by twitching motility test, scanning electron microscope, and microtiter plate assay. Results: It could be revealed that 8/100 isolates were MDR, 90/100 isolates were XDR and pan drug resistance level was shown in only two isolates. 80% of isolates had motile ability through twitching assay, and scan electron microscope study revealed that 76% of XDR isolates could produce different stages of biofilm on coverslip placed in MacConkey broth. Microtiter plate assay revealed 81% of XDR isolates were biofilm producers. Conclusion: it could be concluded that extensive drug resistant P. mirabilis can produce biofilm hence resist several important antibiotics; making treatment of infection among wounded patients is such a challenge in many hospitals in Baghdad.

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Microtiter Plate Assay for Assessment of Listeria monocytogenes Biofilm Formation

Applied and Environmental Microbiology ◽

10.1128/aem.68.6.2950-2958.2002 ◽

2002 ◽

Vol 68 (6) ◽

pp. 2950-2958 ◽

Cited By ~ 529

Author(s):

D. Djordjevic ◽

M. Wiedmann ◽

L. A. McLandsborough

Keyword(s):

Listeria Monocytogenes ◽

Crystal Violet ◽

Biofilm Formation ◽

Microtiter Plate ◽

Cellular Growth ◽

Epifluorescence Microscopy ◽

Simple Method ◽

Microtiter Plate Assay ◽

Plate Assay ◽

Biofilm Production

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

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

Taiwan Veterinary Journal ◽

10.1142/s1682648514500140 ◽

2014 ◽

Vol 40 (02) ◽

pp. 101-107

Author(s):

Min-Tao Wan ◽

Chin-Cheng Chou

Keyword(s):

Bacterial Adhesion ◽

Biofilm Formation ◽

Microtiter Plate ◽

Ecological Niches ◽

Microtiter Plate Assay ◽

Environmental Group ◽

Biofilm Production ◽

The Difference ◽

Wastewater Samples

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

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Biofilm Production and Presence of ica and bap Genes in Staphylococcus aureus Strains Isolated from Cows with Mastitis in the Eastern Poland

Polish Journal of Microbiology ◽

10.33073/pjm-2012-009 ◽

2012 ◽

Vol 61 (1) ◽

pp. 65-69 ◽

Cited By ~ 28

Author(s):

PIOTR SZWEDA ◽

MARTA SCHIELMANN ◽

SŁAWOMIR MILEWSKI ◽

ANETA FRANKOWSKA ◽

ANTONI JAKUBCZAK

Keyword(s):

Biofilm Formation ◽

Protein A ◽

Microtiter Plate ◽

Gene Encoding ◽

Microtiter Plate Assay ◽

Plate Assay ◽

Biofilm Production ◽

Genotypic Analysis ◽

Phenotypic Methods ◽

Encoding Protein

The aim of the study was phenotypic and genotypic analysis of 132 S. aureus strains isolated from mastitis in eastern Poland in respect to their biofilm formation ability. The analysis of the size polymorphism of fragment X in the gene encoding protein A (spa) revealed high genetic differentiation of the analyzed group of isolates. The ability of biofilm formation by the isolates was tested using two phenotypic methods. The Congo Red plate assay was found to be irreproducible and very subjective. More objective results were obtained using the spectrophotometric, microtiter plate assay. Most of the isolates, namely 76/132 (57.6 %) were classified as biofilm producers depending on the value of absorbance in the microtiter plate test. All of the isolates tested were found to possess both icaA and icaD genes, while the bap gene was absent in all strains.

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A Microtiter Plate Assay for the Characterization of Serine Proteases by Their Esterase Activity

Analytical Biochemistry ◽

10.1006/abio.1994.1333 ◽

1994 ◽

Vol 220 (2) ◽

pp. 238-243 ◽

Cited By ~ 21

Author(s):

R.G. Whittaker ◽

M.K. Manthey ◽

D.S. Lebrocque ◽

P.J. Hayes

Keyword(s):

Esterase Activity ◽

Serine Proteases ◽

Microtiter Plate ◽

Microtiter Plate Assay ◽

Plate Assay

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Molecular Characterization of Biofilm-Producing Pseudomonas aeruginosa Isolated from Healthy Pigs and Chicken in India

Indian Journal of Animal Research ◽

10.18805/ijar.b-3903 ◽

2020 ◽

Author(s):

S Chakraborty ◽

T K Dutta ◽

P Roychoudhury ◽

I Samanta ◽

Lalhruaipuii . ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Microtiter Plate ◽

Multidrug Resistant ◽

Identification System ◽

Beta Lactamase ◽

M Gene ◽

Isolation And Identification ◽

Microtiter Plate Assay ◽

Plate Assay

Pseudomonas aeruginosa is considered as the most potent member of multidrug-resistant (MDR) and extremely resistant (XDR) gram-negative pathogens (‘ESCAPE’ group) emerged throughout the world with the property to ‘escape’ the treatment with antibiotics. Biofilm formation is a significant virulence property of P. aeruginosa generating not only antibiotic resistance, but also it acts as a constant source of infection in the host and it can prevent host defence such as chemotaxis of polynuclear immune cells. There is paucity of scientific literatures regarding characterization of biofilm producing P. aeruginosa isolated from livestock and birds. A total of 200 rectal swabs were collected from pigs (n=100) and chickens (n=100) from Mizoram state of India. All the specimens were processed of isolation and identification of P. aeruginosa, which were further confirmed by 16S rRNA PCR and Phoenix bacterial identification system. All the isolates were subjected to detection of biofilm producing ability by microtiter plate assay, antimicrobial sensitivity by disc diffusion assay and detection of selected biofilm producing genes as well as selected beta lactamase genes by specific PCR assay. A total of 11 P. aeruginosa were isolated from pigs (n=6) and chickens (n=5). All the isolates were recorded as positive for biofilm production by microtiter plate assay and were positive for at least one biofilm associated genes. A total of 8 isolates were positive for blaTEM gene and one isolates was positive for blaCTX-M gene. This is probably the unique report on isolation of P. aeruginosa from animals carrying both beta lactamase as well as biofilm producing genes.

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Hydrodynamic Evaluation of Microtiter Plate Assay Using Computational Fluid Dynamics for Biofilm Formation

Volume 2: Computational Fluid Dynamics ◽

10.1115/ajkfluids2019-5425 ◽

2019 ◽

Author(s):

R. Zahra ◽

A. A. Khan ◽

M. Sajid

Keyword(s):

Fluid Dynamics ◽

Computational Fluid Dynamics ◽

Biofilm Formation ◽

Microtiter Plate ◽

Bacterial Cells ◽

Microtiter Plate Assay ◽

Plate Assay ◽

Microtiter Plates ◽

Biofilm Production ◽

Static Conditions

Abstract Biofilms are complex surface associated communities where bacterial cells are enclosed by self-produced extra cellular polymeric substances (EPS), mainly consisting of exopolysaccharides, proteins and extracellular DNA. Treatment of biofilm associated persistent infections is an emerging issue for clinicians as bacterial cells adhere with human epithelial cells or indwelling medical devices such as implants and catheters, used in urinary tract and respiratory infections. Several methods are in practice to assess the biofilm formation of bacterial strains. Most of these are phenotypic methods which include Congo red assay (CRA), Air liquid interface (ALI), tissue culture plate method and Microtiter plate assay (MTPA). MTPA is considered as a standard screening method for comparing adherence pattern and is the most widely used quantitative method for detection of biofilm formation. Generally, the assay is performed under standard static conditions and little is known about the hydrodynamics in the microtiter plates. A few studies have applied computational fluid dynamics (CFD) simulations to describe flow pattern in microtiter plates during biofilm production and optimized the suitable conditions to detect the biofilm formation which have proven to be efficient. In this work the dependencies of biofilm formation on the hydrodynamics in microtiter plate assays were evaluated using OpenFOAM® an open-source toolbox for numerical simulation. It was found that higher flow rates increase the nutrient availability, promote cell growth, and attachment pattern with increased production of exopolymer, while the increase in flow velocity increases the shear rate causing erosion and disassembly of biofilm production because of detachment from the surface.

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Drug Targeting Mycobacterium tuberculosis Cell Wall Synthesis: Development of a Microtiter Plate-Based Screen for UDP-Galactopyranose Mutase and Identification of an Inhibitor from a Uridine-Based Library

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.47.1.378-382.2003 ◽

2003 ◽

Vol 47 (1) ◽

pp. 378-382 ◽

Cited By ~ 55

Author(s):

Michael S. Scherman ◽

Katharine A. Winans ◽

Richard J. Stern ◽

Victoria Jones ◽

Carolyn R. Bertozzi ◽

...

Keyword(s):

Cell Wall ◽

Mycobacterium Tuberculosis ◽

Drug Targeting ◽

Enzyme Inhibitor ◽

Microtiter Plate ◽

Biosynthetic Enzyme ◽

Cell Wall Synthesis ◽

Chemical Library ◽

Microtiter Plate Assay ◽

Plate Assay

ABSTRACT A microtiter plate assay for UDP-galactopyranose mutase, an essential cell wall biosynthetic enzyme of Mycobacterium tuberculosis, was developed. The assay is based on the release of tritiated formaldehyde from UDP-galactofuranose but not UDP-galactopyranose by periodate and was used to identify a uridine-based enzyme inhibitor from a chemical library.

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