scholarly journals Investigation of Biofilm Formation and Phylogenetic Typing of Escherichia Coli Strains Isolated from Milk of Cows with Mastitis / Ispitivanje Formiranja Biofilma I Filogenetska Tipizacija Sojeva Escherichia Coli Izolovanih Iz Mleka Krava Sa Mastitisom

2015 ◽  
Vol 65 (2) ◽  
pp. 202-216 ◽  
Author(s):  
Dubravka Milanov ◽  
Bojana Prunić ◽  
Maja Velhner ◽  
Dalibor Todorović ◽  
Vladimir Polaček
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Mammary Gland ◽  
Antimicrobial Susceptibility ◽  
Biofilm Formation ◽  
Clinical Symptoms ◽  
Disk Diffusion Test ◽  
E Coli ◽  
Extracellular Matrix Components

Abstract Escherichia coli is an opportunistic pathogen affecting bovine mammary gland causing mainly transient infections; however, some recent reports indicated that some strains are able to adhere to and internalize into the epithelial cells, which can result in the persistence of the pathogen in the tissue and development of recurrent mastitis. The mechanism of adaptation of E. coli to the mammary gland relies on structures that are distinctive components of its extracellular matrix - curli fimbriae (proteinaceous component) and cellulose (polysaccharide). Expression of these components varies among the isolates. In this study, we investigated the capacity of expression of curli fimbriae and cellulose (via colony morphotype on Congo Red agar) and ability of biofilm formation (microtiter plate test) in 25 strains of E. coli isolated from milk of cows with clinical mastitis. Phylogenetic grouping of the isolates was performed using PCR method based on detection of chuA, yjaA and TspE4-C2 amplicons. Antimicrobial susceptibility was examined using standard disk diffusion test. Production of both extracellular matrix components was established in 56%, and expression of curli fimbriae in 64% E. coli isolates. All isolates that produced curli fimbriae, demonstrated this ability at a temperature of 37°C, indicating the potential role of these adhesive structures in the pathogenesis of mastitis. The results of phylogenetic typing confirmed that E. coli strains isolated from milk of cows with mastitis are typical commensals mainly belonging to phylogenetic groups A and B1. All curli and curli/cellulose producing isolates formed biofilm under in vitro conditions. The biofilm potentially plays an important role in the development of persistent infections as well as recurrent clinical symptoms after antibiotic therapy in spite of quite good in vitro antimicrobial susceptibility of the agent.

Rosemary and Tea Tree Essential Oils Exert Antibiofilm Activities In Vitro against Staphylococcus aureus and Escherichia coli

2020 ◽  
Vol 83 (7) ◽  
pp. 1261-1267
Author(s):  
TING LIU ◽  
JINGFAN WANG ◽  
XIAOMAN GONG ◽  
XIAOXIA WU ◽  
LIU LIU ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Essential Oil ◽  
Biofilm Formation ◽  
Gas Chromatography Mass Spectrometry ◽  
Tea Tree ◽  
E Coli ◽  
Microdilution Method ◽  
Broth Microdilution Method

ABSTRACT The purpose of the present study was to determine the bioactive compounds in rosemary essential oil (REO) and tea tree essential oil (TEO) and to investigate their antibacterial and antibiofilm activities against Staphylococcus aureus and Escherichia coli in vitro. The MIC and MBC assays were performed to assess the antibacterial activity of these two EOs against S. aureus and E. coli with the broth microdilution method. A crystal violet assay was used to ascertain the effects of EOs on the biofilm formation of the test strains, and a tetrazolium bromide (MTT) assay was used to measure the level of inactivation of mature biofilms by EOs. Gas chromatography–mass spectrometry revealed 15 compounds in REO and 27 compounds in TEO, representing 97.78 and 98.13% of the total EO, respectively. Eucalyptol and α-pinene were found in high concentrations in REO, and the two major compounds in TEO were 4-terpineol and terpinolene. The MICs of REO for the two S. aureus and E. coli test strains were both 0.5 mg/mL, and the MICs of TEO for the two strains were both 0.25 mg/mL. Therefore, these EOs can significantly inhibit the formation of biofilms and induced morphological biofilm changes, as verified by scanning electron microscopy. Both EOs had destructive effects on the mature biofilm of the two test strains. TEO was more inhibitory than REO for biofilm formation by the two test strains. HIGHLIGHTS

scholarly journals The Catabolite Repressor Protein-Cyclic AMP Complex RegulatescsgDand Biofilm Formation in Uropathogenic Escherichia coli

2016 ◽  
Vol 198 (24) ◽  
pp. 3329-3334 ◽  
Author(s):  
David A. Hufnagel ◽  
Margery L. Evans ◽  
Sarah E. Greene ◽  
Jerome S. Pinkner ◽  
Scott J. Hultgren ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Cyclic Amp ◽  
Intergenic Region ◽  
Biofilm Formation ◽  
Cellulose Production ◽  
Content Type ◽  
E Coli ◽  
Link Type ◽  
K1 Capsule

ABSTRACTThe extracellular matrix protectsEscherichia colifrom immune cells, oxidative stress, predation, and other environmental stresses. Production of theE. coliextracellular matrix is regulated by transcription factors that are tuned to environmental conditions. The biofilm master regulator protein CsgD upregulates curli and cellulose, the two major polymers in the extracellular matrix of uropathogenicE. coli(UPEC) biofilms. We found that cyclic AMP (cAMP) regulates curli, cellulose, and UPEC biofilms throughcsgD. The alarmone cAMP is produced by adenylate cyclase (CyaA), and deletion ofcyaAresulted in reduced extracellular matrix production and biofilm formation. Thecataboliterepressorprotein (CRP) positively regulatedcsgDtranscription, leading to curli and cellulose production in the UPEC isolate, UTI89. Glucose, a known inhibitor of CyaA activity, blocked extracellular matrix formation when added to the growth medium. The mutant strains ΔcyaAand Δcrpdid not produce rugose biofilms, pellicles, curli, cellulose, or CsgD. Three putative CRP binding sites were identified within thecsgD-csgBintergenic region, and purified CRP could gel shift thecsgD-csgBintergenic region. Additionally, we found that CRP binded upstream ofkpsMT, which encodes machinery for K1 capsule production. Together our work shows that cAMP and CRP influenceE. colibiofilms through transcriptional regulation ofcsgD.IMPORTANCEThecataboliterepressorprotein (CRP)-cyclic AMP (cAMP) complex influences the transcription of ∼7% of genes on theEscherichia colichromosome (D. Zheng, C. Constantinidou, J. L. Hobman, and S. D. Minchin, Nucleic Acids Res 32:5874–5893, 2004,https://dx.doi.org/10.1093/nar/gkh908). Glucose inhibitsE. colibiofilm formation, and ΔcyaAand Δcrpmutants show impaired biofilm formation (D. W. Jackson, J.W. Simecka, and T. Romeo, J Bacteriol 184:3406–3410, 2002,https://dx.doi.org/10.1128/JB.184.12.3406-3410.2002). We determined that the cAMP-CRP complex regulates curli and cellulose production and the formation of rugose and pellicle biofilms throughcsgD. Additionally, we propose that cAMP may work as a signaling compound for uropathogenicE. coli(UPEC) to transition from the bladder lumen to inside epithelial cells for intracellular bacterial community formation through K1 capsule regulation.

scholarly journals Salmonella Extracellular Matrix Components Influence Biofilm Formation and Gallbladder Colonization

2016 ◽  
Vol 84 (11) ◽  
pp. 3243-3251 ◽  
Author(s):  
Haley E. Adcox ◽  
Erin M. Vasicek ◽  
Varun Dwivedi ◽  
Ky V. Hoang ◽  
Joanne Turner ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Typhoid Fever ◽  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Bacterial Survival ◽  
Content Type ◽  
Extracellular Matrix Components ◽  
Matrix Components

Salmonella enterica serovar Typhi, the causative agent of typhoid fever in humans, forms biofilms encapsulated by an extracellular matrix (ECM). Biofilms facilitate colonization and persistent infection in gallbladders of humans and mouse models of chronic carriage. Individual roles of matrix components have not been completely elucidated in vitro or in vivo . To examine individual functions, strains of Salmonella enterica serovar Typhimurium, the murine model of S . Typhi, in which various ECM genes were deleted or added, were created to examine biofilm formation, colonization, and persistence in the gallbladder. Studies show that curli contributes most significantly to biofilm formation. Expression of Vi antigen decreased biofilm formation in vitro and virulence and bacterial survival in vivo without altering the examined gallbladder pro- or anti-inflammatory cytokines. Oppositely, loss of all ECM components (Δ wcaM Δ csgA Δ yihO Δ bcsE ) increased virulence and bacterial survival in vivo and reduced gallbladder interleukin-10 (IL-10) levels. Colanic acid and curli mutants had the largest defects in biofilm-forming ability and contributed most significantly to the virulence increase of the Δ wcaM Δ csgA Δ yihO Δ bcsE mutant strain. While the Δ wcaM Δ csgA Δ yihO Δ bcsE mutant was not altered in resistance to complement or growth in macrophages, it attached and invaded macrophages better than the wild-type (WT) strain. These data suggest that ECM components have various levels of importance in biofilm formation and gallbladder colonization and that the ECM diminishes disseminated disease in our model, perhaps by reducing cell attachment/invasion and dampening inflammation by maintaining/inducing IL-10 production. Understanding how ECM components aid acute disease and persistence could lead to improvements in therapeutic treatment of typhoid fever patients.

scholarly journals In VitroAntimicrobial Resistance of Urinary Escherichia coli Isolates among U.S. Outpatients from 2000 to 2010

2012 ◽  
Vol 56 (4) ◽  
pp. 2181-2183 ◽  
Author(s):  
Guillermo V. Sanchez ◽  
Ronald N. Master ◽  
James A. Karlowsky ◽  
Jose M. Bordon
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Antimicrobial Susceptibility ◽  
Minimal Change ◽  
Urine Samples ◽  
Surveillance Network ◽  
Content Type ◽  
E Coli ◽  
Resistance Data

ABSTRACTThis study examinesin vitroantimicrobial resistance data fromEscherichia coliisolates obtained from urine samples of U.S. outpatients between 2000 and 2010 using The Surveillance Network (TSN). Antimicrobial susceptibility results (n= 12,253,679) showed the greatest increases inE. coliresistance from 2000 to 2010 for ciprofloxacin (3% to 17.1%) and trimethoprim-sulfamethoxazole (TMP-SMX) (17.9% to 24.2%), whereas nitrofurantoin (0.8% to 1.6%) and ceftriaxone (0.2% to 2.3%) showed minimal change. From 2000 to 2010, the antimicrobial resistance of urinaryE. coliisolates to ciprofloxacin and TMP-SMX among outpatients increased substantially.

scholarly journals Differential Binding of Escherichia coli O157:H7 to Alfalfa, Human Epithelial Cells, and Plastic Is Mediated by a Variety of Surface Structures

2005 ◽  
Vol 71 (12) ◽  
pp. 8008-8015 ◽  
Author(s):  
Alfredo G. Torres ◽  
Cecelia Jeter ◽  
William Langley ◽  
Ann G. Matthysse
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Food Poisoning ◽  
The United States ◽  
Surface Proteins ◽  
E Coli ◽  
Alfalfa Sprouts ◽  
K 12 ◽  
Plant Surfaces

ABSTRACT Escherichia coli O157:H7 carried on plant surfaces, including alfalfa sprouts, has been implicated in food poisoning and outbreaks of disease in the United States. Adhesion to cell surfaces is a key component for bacterial establishment and colonization on many types of surfaces. Several E. coli O157:H7 surface proteins are thought to be important for adhesion and/or biofilm formation. Therefore, we examined whether mutations in several genes encoding potential adhesins and regulators of adherence have an effect on bacterial binding to plants and also examined the role of these genes during adhesion to Caco-2 cells and during biofilm formation on plastic in vitro. The genes tested included those encoding adhesins (cah, aidA1, and ompA) and mediators of hyperadherence (tdcA, yidE, waaI, and cadA) and those associated with fimbria formation (csgA, csgD, and lpfD2). The introduction of some of these genes (cah, aidA1, and csg loci) into an E. coli K-12 strain markedly increased its ability to bind to alfalfa sprouts and seed coats. The addition of more than one of these genes did not show an additive effect. In contrast, deletion of one or more of these genes in a strain of E. coli O157:H7 did not affect its ability to bind to alfalfa. Only the absence of the ompA gene had a significant effect on binding, and the plant-bacterium interaction was markedly reduced in a tdcA ompA double mutant. In contrast, the E. coli O157:H7 ompA and tdcA ompA mutant strains were only slightly affected in adhesion to Caco-2 cells and during biofilm formation. These findings suggest that some adhesins alone are sufficient to promote binding to alfalfa and that they may exist in E. coli O157:H7 as redundant systems, allowing it to compensate for the loss of one or more of these systems. Binding to the three types of surfaces appeared to be mediated by overlapping but distinct sets of genes. The only gene which appeared to be irreplaceable for binding to plant surfaces was ompA.

scholarly journals Bacterial Profile of Urinary Tract Infections: Evaluation of Biofilm Formation and Antibiotic Resistance Pattern of Uropathogenic Escherichia coli

2020 ◽  
Vol 14 (4) ◽  
pp. 2577-2584
Author(s):  
Tariq Ahmad Shah ◽  
P. Preethishree ◽  
Ashwini ◽  
Vidya Pai
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Biofilm Formation ◽  
Urine Samples ◽  
Diffusion Method ◽  
E Coli ◽  
Biofilm Production ◽  
Tract Infections

Urinary tract infection (UTI) is one of the most common complaints in the outpatient clinic and a major health problem owing to the emergence of antibiotic resistance and biofilm formation. The objective of this study was to isolate and identify the causative bacterial agent of UTI and detect in vitro biofilm formation by Escherichia coli and investigate its correlation with antibiotic resistance. Urine samples from 519 patients with suspected UTIs were collected and processed by conventional microbiological procedures. Antimicrobial susceptibility testing for E. coli isolates was performed on Mueller Hinton agar (MHA) plates using the Kirby-Bauer disk diffusion method. Biofilm production was evaluated using the tissue culture plate method. Of 519 urine samples, 115 (22.1%) showed significant bacteriuria. The most common isolate was E. coli (n=57, 49.6%), followed by Klebsiella spp. (n=23, 20%). All E. coli isolates were evaluated for their ability to form biofilms in vitro. Of 57 isolates, 50 (87.7%) were biofilm producers and 7 (12.3%) were non-biofilm producers. Antibiogram of E. coli isolates revealed the highest resistance to ampicillin (96.5%) and nitrofurantoin (91.2%), followed by amoxyclav (82.5%), ceftazidime (73.7%), cefepime (71.9%), and tetracycline (71.9%). A significant association (p<0.05) was observed between biofilm formation and resistance to amoxyclav, ceftazidime, cefepime, imipenem, and nitrofurantoin. A significant correlation was noted between biofilm production and antibiotic resistance. Hence, screening of all isolates of uropathogenic E. coli for biofilm production and studying their antibiogram would allow appropriate choice of antibiotic therapy.

scholarly journals Determination of Extended-Spectrum β-Lactamases and AmpC Production in Uropathogenic Isolates of Escherichia coli and Susceptibility to Fosfomycin

2013 ◽  
Vol 5 (02) ◽  
pp. 090-093 ◽  
Author(s):  
Varsha Gupta ◽  
Hena Rani ◽  
Nidhi Singla ◽  
Neelam Kaistha ◽  
Jagdish Chander
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Susceptibility ◽  
Boronic Acid ◽  
Treatment Options ◽  
Community Setting ◽  
Integrated Approach ◽  
Susceptibility Pattern ◽  
E Coli ◽  
Antimicrobial Susceptibility Pattern

ABSTRACT Background: Urinary tract infection due to Escherichia coli is one of the common problem in clinical practice. Various drug resistance mechanisms are making the bacteria resistant to higher group of drugs making the treatment options very limited. This study was undertaken to detect ESBLs and AmpC production in uropathogenic Escherichia coli isolates and to determine their antimicrobial susceptibility pattern with special reference to fosfomycin. Materials and Methods: A total number of 150 E. coli isolates were studied. ESBL detection was done by double disc synergy and CLSI method. AmpC screening was done using cefoxitin disc and confirmation was done using cefoxitin/cefoxitin-boronic acid discs. In AmpC positive isolates, ESBLs was detected by modifying CLSI method using boronic acid. Antimicrobial susceptibility pattern was determined following CLSI guidelines. Fosfomycin susceptibility was determined by disc diffusion and E-test methods. Results: ESBLs production was seen in 52.6% of isolates and AmpC production was seen in 8% of isolates. All AmpC producers were also found to be ESBLs positive. ESBLs positive isolates were found to be more drug resistant than ESBLs negative isolates. All the strains were found to be fosfomycin sensitive. Conclusions: ESBLs and AmpC producing isolates are becoming prevalent in E. coli isolates from community setting also. Amongst the oral drugs, no in-vitro resistance has been seen for fosfomycin making it a newer choice of drug (although not new) in future. An integrated approach to contain antimicrobial resistance should be actually the goal of present times.

scholarly journals Genome rearrangements induce biofilm formation in Escherichia coli C – an old model organism with a new application in biofilm research

BMC Genomics ◽  
2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Jarosław E. Król ◽  
Donald C. Hall ◽  
Sergey Balashov ◽  
Steven Pastor ◽  
Justin Sibert ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Insertion Sequence ◽  
Genomic Sequence ◽  
Model Organism ◽  
Start Codon ◽  
Bacterial Survival ◽  
E Coli ◽  
Full Genomic Sequence

Abstract Background Escherichia coli C forms more robust biofilms than other laboratory strains. Biofilm formation and cell aggregation under a high shear force depend on temperature and salt concentrations. It is the last of five E. coli strains (C, K12, B, W, Crooks) designated as safe for laboratory purposes whose genome has not been sequenced. Results Here we present the complete genomic sequence of this strain in which we utilized both long-read PacBio-based sequencing and high resolution optical mapping to confirm a large inversion in comparison to the other laboratory strains. Notably, DNA sequence comparison revealed the absence of several genes thought to be involved in biofilm formation, including antigen 43, waaSBOJYZUL for lipopolysaccharide (LPS) synthesis, and cpsB for curli synthesis. The first main difference we identified that likely affects biofilm formation is the presence of an IS3-like insertion sequence in front of the carbon storage regulator csrA gene. This insertion is located 86 bp upstream of the csrA start codon inside the − 35 region of P4 promoter and blocks the transcription from the sigma32 and sigma70 promoters P1-P3 located further upstream. The second is the presence of an IS5/IS1182 in front of the csgD gene. And finally, E. coli C encodes an additional sigma70 subunit driven by the same IS3-like insertion sequence. Promoter analyses using GFP gene fusions provided insights into understanding this regulatory pathway in E. coli. Conclusions Biofilms are crucial for bacterial survival, adaptation, and dissemination in natural, industrial, and medical environments. Most laboratory strains of E. coli grown for decades in vitro have evolved and lost their ability to form biofilm, while environmental isolates that can cause infections and diseases are not safe to work with. Here, we show that the historic laboratory strain of E. coli C produces a robust biofilm and can be used as a model organism for multicellular bacterial research. Furthermore, we ascertained the full genomic sequence of this classic strain, which provides for a base level of characterization and makes it useful for many biofilm-based applications.

Sign in / Sign up

Export Citation Format

Share Document