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

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

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

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

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

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

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

2020 ◽  
Vol 11 (2) ◽  
pp. 70-78
Author(s):  
Rawa Aziz ◽  
◽  
Elaf Mohameed
Keyword(s):  
Electron Microscope ◽  
Antibiotic Sensitivity ◽  
Microtiter Plate ◽  
Automated System ◽  
Drug Resistant ◽  
Resistance Level ◽  
Microtiter Plate Assay ◽  
Plate Assay ◽  
Biofilm Production

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.

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

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

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

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

2012 ◽  
Vol 61 (1) ◽  
pp. 65-69 ◽  
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.

scholarly journals Comparative Characterisation of Genotypically Different Clones of MRSA in the Production of Biofilms

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Salman Sahab Atshan ◽  
Mariana Nor Shamsudin ◽  
Leslie Than Thian Lung ◽  
Zamberi Sekawi ◽  
Ehsanollah Ghaznavi-Rad ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Protein A ◽  
Surface Protein ◽  
Microtiter Plate ◽  
Microtiter Plate Assay ◽  
Biofilm Production ◽  
Production Capacities ◽  
Glass Surfaces ◽  
Sccmec Types ◽  
Spa Types

The ability to adhere and produce biofilms is characteristic of enhanced virulence among isolates of methicillin-resistantStaphylococcus aureus(MRSA). The aim of the study is to find out whether these characteristics are consistently similar among isolates variations of MRSA. The study used 30 various isolates of MRSA belong to 13 spa types and 5 MLST types and determined the aggregation, the adherence, and the production of biofilms and slime for each isolate. The methods used to evaluate these characteristics were a modified Congo red agar assay (MCRA), a microtiter plate assay (MPA), high-magnification light microscopy, scanning electron microscopy (SEM), and PCR. The study found that isolates belonging to similar Spa, SCCmec, and ST types have similar abilities to produce biofilms; however, their ability to produce slime on CRA was found to be different. Moreover, isolates that have different Spa types showed high variation in their ability to produce biofilms. The results of light microscope revealed the isolates that produced strong and weak biofilms and formed similar aggregation on the glass surfaces. SEM results showed that all 30 MRSA isolates that were tested were 100% positive for biofilm formation, although to varying degrees. Further testing using PCR confirmed that 100% of the 30 isolates tested were positive for the presence of theicaADBC,fnbA, eno, ebps, clfA, andclfBgenes. The prevalence offib, cna, fnbB, andbbpin MRSA clones was 90, 93.33, 53.33, and 10%, respectively. This study indicate that differences in biofilm production capacities are caused by the differences in surface protein A (Spa) type and are not due to differences in MLST and SCCmec types.

scholarly journals Analysis of the bacterial biofilm formation in different models of the in vitro culture

2021 ◽  
Vol 19 (1) ◽  
pp. 40-45
Author(s):  
Agnieszka Bogut ◽  
◽  
Agnieszka Magryś ◽  
Keyword(s):  
Biofilm Formation ◽  
Culture Media ◽  
Microtiter Plate ◽  
Bacterial Biofilm ◽  
Atmospheric Conditions ◽  
Experimental Conditions ◽  
Microtiter Plate Assay ◽  
Biofilm Production ◽  
Strain Type

Introduction. Microtiter plate assay (MPA) remains one of workhorses of in vitro biofilm research but it requires optimization of experimental conditions to fulfill the biofilm formation requirements of different bacterial pathogens. Aim. The aim was to determine the effect of TSB and RPMI1640 culture media and selected culture variables (O2 vs. 5% CO2, extended incubation time) on the biofilm production by bacteria commonly involved in biofilm-related infections: Enterococcus faecalis (EF), Escherichia coli (EC), Staphylococcus aureus (SA), Pseudomonas aeruginosa (PA), Klebsiella pneumoniae (KP). Material and methods. The investigation was performed using the MPA with crystal violet. Results. Statistically significant (p<0.05) increase in biofilm production between 24h and 72h time points was observed for EF (TSB o2, RPMIo2 and RPMIco2), EC (TSBo2), SA (TSBo2, TSBco2), KP (TSBo2, TSBco2), PA (RPMIco2, TSBco2). The TSB caused a significantly greater stimulation of biofilm production compared to RPM1640. It outcompeted RPMI1640 irrespective of the atmospheric conditions for SA and KP and under aerobic conditions for EF. Conclusion. Although the TSB provided the most optimal conditions for biofilm production, the process was influenced by the strain type, atmospheric conditions and period of cultivation which limits the ability to design a single universal model of the in vitro biofilm investigation.

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

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

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

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

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.

scholarly journals Association of Oral Microbiota Properties on Hyaluronidase and Biofilm Formation

2021 ◽  
Vol 15 ◽  
pp. 1-7
Author(s):  
Nur Syarafina Mohd Zahir ◽  
Nabilah Huda Abdul Halim ◽  
Hanani Ahmad Yusof
Keyword(s):  
Biofilm Formation ◽  
Bacterial Species ◽  
Microtiter Plate ◽  
Oral Microbiota ◽  
Gram Negative Bacteria ◽  
Microtiter Plate Assay ◽  
Gram Staining ◽  
Biofilm Production ◽  
Biofilm Detachment ◽  
Colony Surface

Correlation between hyaluronidase (Hyl) activity and biofilm detachment in a few bacterial species was found. However, it is unclear if this association applies to bacterial species or for more general bacterial characteristics. This study determined the association between biofilm production and Hyl activity among bacterial isolates from the oral cavity of healthy subjects, and its association with Gram staining group, colony surface morphology and bacteria shape.  The swab was taken from the tongue, cheek and entire teeth surfaces of 35 subjects, and tested for biofilm through modified microtiter plate assay while Hyl production was screened through HA rapid plate method. Forty-four isolates were found, each 50% are Gram-positive, and Gram-negative bacteria, with the majority are cocci and non-mucoid colony. More than 70% of isolates are moderate and strong; (n= 17, 38.6%) and (n=15, 34.1%) respectively for biofilm production; and 68.2% are Hyl producer. A significant association was found between Hyl and bacterial shape (p=0.018) and colony morphology (p=0.018), while other association is not significantly measured, including between Hyl and biofilm (p=0.659). This study showed that biofilm production is not affected by the characteristics of the bacteria to produce or not produce hyaluronidase. Meanwhile, Hyl production is prone in rod shape and mucoid isolates which need further investigations.

scholarly journals Biofilm formation capacity of Salmonella serotypes at different temperature conditions

2018 ◽  
Vol 38 (1) ◽  
pp. 71-76 ◽  
Author(s):  
Karen A. Borges ◽  
Thales Q. Furian ◽  
Sara N. Souza ◽  
Rafaela Menezes ◽  
Eduardo C. Tondo ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Production Capacity ◽  
Microtiter Plate ◽  
Bacterial Cells ◽  
Bacterial Survival ◽  
Salmonella Spp ◽  
Microtiter Plate Assay ◽  
Temperature Conditions ◽  
Biofilm Production ◽  
Salmonella Serotypes

ABSTRACT: Salmonella spp. are one of the most important agents of foodborne disease in several countries, including Brazil. Poultry-derived products are the most common food products, including meat and eggs, involved in outbreaks of human salmonellosis. Salmonella has the capacity to form biofilms on both biotic and abiotic surfaces. The biofilm formation process depends on an interaction among bacterial cells, the attachment surface and environmental conditions. These structures favor bacterial survival in hostile environments, such as slaughterhouses and food processing plants. Biofilms are also a major problem for public health because breakage of these structures can cause the release of pathogenic microorganisms and, consequently, product contamination. The aim of this study was to determine the biofilm production capacity of Salmonella serotypes at four different temperatures of incubation. Salmonella strains belonging to 11 different serotypes, isolated from poultry or from food involved in salmonellosis outbreaks, were selected for this study. Biofilm formation was investigated under different temperature conditions (37°, 28°, 12° and 3°C) using a microtiter plate assay. The tested temperatures are important for the Salmonella life cycle and to the poultry-products process. A total of 92.2% of the analyzed strains were able to produce biofilm on at least one of the tested temperatures. In the testing, 71.6% of the strains produced biofilm at 37°C, 63% at 28°C, 52.3% at 12°C and 39.5% at 3°C, regardless of the serotype. The results indicate that there is a strong influence of temperature on biofilm production, especially for some serotypes, such as S. Enteritidis, S. Hadar and S. Heidelberg. The production of these structures is partially associated with serotype. There were also significant differences within strains of the same serotype, indicating that biofilm production capacity may be strain-dependent.

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