Association between biofilm-production and antibiotic resistance in Escherichia coli isolates: A laboratory-based case study and a literature review

AbstractBacteria can enhance their survival by attaching to inanimate surfaces or tissues, and presenting as multicellular communities encased in a protective extracellular matrix called biofilm. There has been pronounced interest in assessing the relationship between the antibiotic resistant phenotype and biofilm-production in clinically-relevant pathogens. The aim of the present paper was to provide additional experimental results on the topic, testing the biofilm-forming capacity of Escherichia coli isolates using in vitro methods in the context of their antibiotic resistance in the form of a laboratory case study, in addition to provide a comprehensive review of the subject. In our case study, a total of two hundred and fifty (n = 250) E. coli isolates, originating from either clean-catch urine samples (n = 125) or invasive samples (n = 125) were included. The colony morphology of isolates were recorded after 24h, while antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method. Biofilm-formation of the isolates was assessed with the crystal violet tube-adherence method. Altogether 57 isolates (22.8%) isolates were multidrug resistant (MDR), 89 isolates (35.6%) produced large colonies (>3 mm), mucoid variant colonies were produced in 131 cases (52.4%), and 108 (43.2%) were positive for biofilm formation. Biofilm-producers were less common among isolates resistant to third-generation cephalosporins and trimethoprim-sulfamethoxazole (P = 0.043 and P = 0.023, respectively). Biofilms facilitate a protective growth strategy in bacteria, ensuring safety against environmental stressors, components of the immune system and noxious chemical agents. Being an integral part of bacterial physiology, biofilm-formation is interdependent with the expression of other virulence factors (especially adhesins) and quorum sensing signal molecules. More research is required to allow for the full understanding of the interplay between the MDR phenotype and biofilm-production, which will facilitate the development of novel therapeutic strategies.

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Association between Biofilm-Production and Antibiotic Resistance in Uropathogenic Escherichia coli (UPEC): An In Vitro Study

Diseases ◽

10.3390/diseases8020017 ◽

2020 ◽

Vol 8 (2) ◽

pp. 17 ◽

Cited By ~ 6

Author(s):

Payam Behzadi ◽

Edit Urbán ◽

Márió Gajdács

Keyword(s):

Escherichia Coli ◽

Biofilm Formation ◽

In Vitro Study ◽

Point Of View ◽

Diffusion Method ◽

Medical Attention ◽

Disk Diffusion Method ◽

Biofilm Production ◽

Tract Infections

Urinary tract infections (UTIs) are among the most common infections requiring medical attention worldwide. The production of biofilms is an important step in UTIs, not only from a mechanistic point of view, but this may also confer additional resistance, distinct from other aspects of multidrug resistance (MDR). A total of two hundred and fifty (n = 250) Escherichia coli isolates, originating from clean-catch urine samples, were included in this study. The isolates were classified into five groups: wild-type, ciprofloxacin-resistant, fosfomycin-resistant, trimethoprim-sulfamethoxazole-resistant and extended spectrum β-lactamase (ESBL)-producing strains. The bacterial specimens were cultured using eosine methylene blue agar and the colony morphology of isolates were recorded. Antimicrobial susceptibility testing was performed using the Kirby–Bauer disk diffusion method and E-tests. Biofilm-formation of the isolates was carried out with the crystal violet tube-adherence method. n = 76 isolates (30.4%) produced large colonies (>3 mm), mucoid variant colonies were produced in n = 135 cases (54.0%), and n = 119 (47.6%) were positive for biofilm formation. The agreement (i.e., predictive value) of mucoid variant colonies in regard to biofilm production in the tube-adherence assay was 0.881 overall. Significant variation was seen in the case of the group of ESBL-producers in the ratio of biofilm-producing isolates. The relationship between biofilm-production and other resistance determinants has been extensively studied. However, no definite conclusion can be reached from the currently available data.

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

Journal of Pure and Applied Microbiology ◽

10.22207/jpam.14.4.33 ◽

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.

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The Frequency of Fluoroquinolone Resistant Enterotoxigenic Escherichia coli from Stools of Diarrheic Children in Ebonyi State, Nigeria

10.21203/rs.2.10616/v1 ◽

2019 ◽

Author(s):

Ebuka David ◽

MA Yameen ◽

IO Igwenyi ◽

IR Iroha ◽

HC Nzelibe ◽

...

Keyword(s):

Escherichia Coli ◽

Biofilm Formation ◽

Enterotoxigenic Escherichia Coli ◽

Fluoroquinolone Resistance ◽

Diffusion Method ◽

Drug Resistant ◽

Disk Diffusion Method ◽

E Coli ◽

Biofilm Production ◽

Rapid Emergence

Abstract Enterotoxigenic Escherichia coli (ETEC) is an important cause of acute childhood diarrhea. The evaluation of ETEC in children is important for therapeutic and economic purposes. Hence, this study aimed to evaluate the frequency of ETEC among diarrheic children, their multidrug and fluoroquinolone resistant pattern. A total of twenty diarrheagenic E. coli (DEC) isolates were gotten from hundred diarrheal samples using biochemical and molecular methods. Multiplex PCR was used to detect the presence of four different pathological types of DEC. Disk diffusion method was used to determine the antibiotic susceptibility of the organisms. Biofilm formation was detected by thiazoylblue tetrazolium bromide dye in a 96-well plate. Results showed that ETEC represented 30% of the DEC, of which 80% were multidrug and fluoroquinolone resistant. The biofilm production abilities of all the ETEC were found to exist within weak, moderate and strong biofilm producers. We observed a high ETEC frequency and rapid emergence of multidrug/fluoroquinolone resistance, suggesting that it is one of the most important causes of frequent drug resistant diarrhea in children in this region.

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A Preliminary Study: Antibiotic Resistance Patterns of Escherichia coli and Enterococcus Species from Wildlife Species Subjected to Supplementary Feeding on Various South African Farms

Animals ◽

10.3390/ani10030396 ◽

2020 ◽

Vol 10 (3) ◽

pp. 396 ◽

Cited By ~ 1

Author(s):

Michaela Sannettha van den Honert ◽

Pieter Andries Gouws ◽

Louwrens Christiaan Hoffman

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Supplementary Feeding ◽

Health Concern ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Wildlife Species ◽

Human Contact ◽

Resistance Patterns

Studies have shown that antibiotic resistance among wild animals is becoming a public health concern, owing to increased contact and co-habitation with domestic animals that, in turn, results in increased human contact, indirectly and directly. This type of farming practice intensifies the likelihood of antibiotic resistant traits in microorganisms transferring between ecosystems which are linked via various transfer vectors, such as rivers and birds. This study aimed to determine whether the practice of wildlife supplementary feeding could have an influence on the antibiotic resistance of the bacteria harboured by the supplementary fed wildlife, and thus play a potential role in the dissemination of antibiotic resistance throughout nature. Escherichia coli and Enterococcus were isolated from the faeces of various wildlife species from seven different farms across South Africa. The Kirby-Bauer disk diffusion method was used according to the Clinical and Laboratory Standards Institute 2018 guidelines. The E. coli (F: 57%; N = 75% susceptible) and Enterococcus (F: 67%; N = 78% susceptible) isolates from the supplementary fed (F) wildlife were in general, found to be more frequently resistant to the selection of antibiotics than from those which were not supplementary fed (N), particularly towards tetracycline (E. coli F: 56%; N: 71%/Enterococcus F: 53%; N: 89% susceptible), ampicillin (F: 82%; N = 95% susceptible) and sulphafurazole (F: 68%; N = 98% susceptible). Interestingly, high resistance towards streptomycin was observed in the bacteria from both the supplementary fed (7% susceptible) and non-supplementary fed (6% susceptible) wildlife isolates. No resistance was found towards chloramphenicol and ceftazidime.

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Detection of biofilm among uropathogenic Escherichia coli and its correlation with antibiotic resistance pattern

Journal of Laboratory Physicians ◽

10.4103/jlp.jlp_98_18 ◽

2019 ◽

Vol 11 (01) ◽

pp. 017-022 ◽

Cited By ~ 5

Author(s):

Rashmi M. Karigoudar ◽

Mahesh H. Karigoudar ◽

Sanjay M. Wavare ◽

Smita S. Mangalgi

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Biofilm Formation ◽

Agar Plate ◽

Diffusion Method ◽

Resistance Pattern ◽

Susceptibility Pattern ◽

Sensitivity Testing ◽

E Coli ◽

Tract Infections

Abstract BACKGROUND: Escherichia coli accounts for 70%–95% of urinary tract infections (UTIs). UTI is a serious health problem with respect to antibiotic resistance and biofilms formation being the prime cause for the antibiotic resistance. Biofilm can restrict the diffusion of substances and binding of antimicrobials. In this context, the present study is aimed to perform in vitro detection of biofilm formation among E. coli strains isolated from urine and to correlate their susceptibility pattern with biofilm formation. MATERIALS AND METHODS: A total of 100 E. coli strains isolated from patients suffering from UTI were included in the study. The identification of E. coli was performed by colony morphology, Gram staining, and standard biochemical tests. The detection of biofilm was carried out by Congo Red Agar (CRA) method, tube method (TM), and tissue culture plate (TCP) method. Antimicrobial sensitivity testing was performed by Kirby–Bauer disc diffusion method on Muller–Hinton agar plate. RESULTS: Of the 100 E. coli strains, 49 (49%) and 51 (51%) were from catheterized and noncatheterized patients, respectively. Biofilm production was positive by CRA, TM, and TCP method were 49 (49%), 55 (55%), and 69 (69%), respectively. Biofilm producers showed maximum resistance to co-trimoxazole (73.9%), gentamicin (94.2%), and imipenem (11.6%) when compared to nonbiofilm producers. Significant association was seen between resistance to antibiotic and biofilm formation with a P = 0.01 (<0.05). CONCLUSION: A greater understanding of biofilm detection in E. coli will help in the development of newer and more effective treatment. The detection of biofilm formation and antibiotic susceptibility pattern helps in choosing the correct antibiotic therapy.

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Antibiotic resistance and virulence genes of extraintestinal pathogenic Escherichia coli from tropical estuary, south India

The Journal of Infection in Developing Countries ◽

10.3855/jidc.5627 ◽

2015 ◽

Vol 9 (05) ◽

pp. 496-504 ◽

Cited By ~ 1

Author(s):

Divya Sukumaran ◽

Abdulla A Mohamed Hatha

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Multidrug Resistance ◽

Virulence Factor ◽

Virulence Genes ◽

Antimicrobial Agents ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli ◽

Virulence Factor Genes

Introduction: Escherichia coli strains can cause a variety of intestinal and extraintestinal diseases. Extraintestinal pathogenic E. coli (ExPEC) strains have the ability to cause severe extraintestinal infections. Multidrug resistance among ExPEC could complicate human infections. Methodology: Escherichia coli strains were isolated during the period of January 2010 to December 2012 from five different stations set at Cochin estuary. Susceptibility testing was determined by the disk-diffusion method using nine different antimicrobial agents. A total of 155 strains of Escherichia coli were screened for the presence of virulence factor genes including papAH, papC, sfa/focDE, iutA,and kpsMT II associated with ExPEC. Results: Among the 155 E. coli isolates, 26 (16.77%), carried two or more virulence genes typical of ExPEC. Furthermore, 19.23% of the ExPEC isolates with multidrug resistance were identified to belong to phylogenetic groups B2 and D. Statistically significant association of iutA gene in ExPEC was found with papC (p < 0.001) and kpsMT II (p < 0.001) genes. ExPEC isolates were mainly resistant to ampicillin (23.07%), tetracycline (19.23%), co-trimoxazole (15.38%), and cefotaxime (15.38%). The adhesion genes papAH and sfa/focDE were positively associated with resistance to gentamicin, chloramphenicol, and cefotaxime (p < 0.05). Conclusions: Co-occurrence of virulence factor genes with antibiotic resistance among ExPEC poses considerable threat to those who use this aquatic system for a living and for recreation.

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Investigation of Antibiotic Resistance and Biofilm Formation in Clinical Isolates of Klebsiella pneumoniae

International Journal of Microbiology ◽

10.1155/2021/5573388 ◽

2021 ◽

Vol 2021 ◽

pp. 1-6

Author(s):

Kiana Karimi ◽

Omid Zarei ◽

Parinaz Sedighi ◽

Mohammad Taheri ◽

Amin Doosti-Irani ◽

...

Keyword(s):

Antibiotic Resistance ◽

Klebsiella Pneumoniae ◽

Biofilm Formation ◽

Diffusion Method ◽

Clinical Samples ◽

Resistance Pattern ◽

Chi Square ◽

Disk Diffusion Method ◽

High Level ◽

Tract Infections

Aim. Klebsiella pneumoniae (K. pneumoniae) is an encapsulated Gram-negative bacterium that can lead to 14–20% of nosocomial infections. The ability of biofilm formation in this bacterium decreases the host immune response and antibiotic efficacy. This may impose a huge impact on patients and healthcare settings. This study aimed to evaluate the antibiotic resistance pattern and biofilm formation in K. pneumoniae strains isolated from two major Hamadan hospitals, west of Iran. Methods. A total of 83 K. pneumoniae strains were isolated from clinical samples of patients in different wards of Hamadan hospitals from September 2018 to March 2019. Determination of antimicrobial susceptibility was performed using the disk diffusion method. Biofilm formation was evaluated by the crystal violet method. Data were analyzed by the SPSS software and chi-square test. Results. The results showed that clinical samples included 18 urinary tract samples (22%), 6 wound samples (7%), 6 blood samples (7%), 17 tracheal tube aspiration samples (20%), 32 throat cultures (38%), 2 sputum samples (2.5%), and 2 abscess drain cultures (2.5%). High-level resistance to cefotaxime was detected in 92%, and all of isolates were susceptible to colistin. Biofilm formation was seen in 62 (75%) isolates. Strong biofilm formation was observed in 17 (20%) strains. A significant correlation was seen between biofilm formation and antibiotic resistance ( P value <0.05). Conclusion. Our findings emphasize the need for proper diagnosis, control, and treatment of infections caused by K. pneumoniae especially in respiratory tract infections due to the strong biofilm formation and high antibiotic resistance in these strains.

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Prevalence of class 1 and 2 integrons among the multidrug resistant uropathogenic strains of Escherichia coli

Asian Biomedicine ◽

10.5372/1905-7415.0901.367 ◽

2017 ◽

Vol 9 (1) ◽

pp. 49-54 ◽

Cited By ~ 1

Author(s):

Haddadi Azam ◽

Somayeh Mikaili Ghezeljeh ◽

Shavandi Mahmoud

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Resistance Genes ◽

Antibiotic Resistance Genes ◽

Multidrug Resistant ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Integrase Gene ◽

Class 1 ◽

Tract Infections

Abstract Background Multidrug resistance is a serious problem in the treatment of urinary tract infections. Horizontal gene transfer, directed by strong selective pressure of antibiotics, has resulted in the widespread distribution of multiple antibiotic resistance genes. The dissemination of resistance genes is enhanced when they are trapped in integrons. Objectives To determine the prevalence of integrons among multidrug resistant Escherichia coli strains collected from regional hospitals and private clinical laboratories in Alborz province. Methods The susceptibility of 111 clinical Escherichia coli isolates was tested using a Kirby–Bauer disk diffusion method for common antibiotics. Isolates were screened for the production of extended spectrum β-lactamases (ESBLs) using a double disk synergy test. The existence of integrons was confirmed by amplification of the integrase gene and their class determined via analysis of PCR products by PCR-RFLP. Results Isolates showed the highest resistance to amoxicillin. Nitrofurantoin, amikacin, and ceftizoxime were the most effective antibiotics in vitro. Eighty-eight isolates of 111 (79%) were resistant to more than three unrelated drugs. We found 30% of the multidrug resistant isolates harbor integrons. Class 1 and 2 integrons were detected in 25 and 1 isolates, respectively. ESBL screening of strains showed 45 isolates (40%) were positive; 22% of the ESBL-positive isolates carried class 1 integrons and the frequency of MDR in ESBLpositive isolates was 93%. Conclusion The existence of integrons in only 29.5% of multidrug resistant isolates showed that besides integrons, antibiotic resistance genes were probably carried on other transferable elements lacking integrons, such as transposons or plasmids.

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Antimicrobial Resistance in Uropathogenic Escherichia coli Strains Isolated from Beasat Hospital in Sanandaj, West of Iran

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2020/v32i130392 ◽

2020 ◽

pp. 32-36

Author(s):

Nasrin Bahmani ◽

Noshin Abdolmaleki ◽

Afshin Bahmani

Keyword(s):

Escherichia Coli ◽

Urinary Tract Infection ◽

Antibiotic Resistance ◽

Urinary Tract ◽

Tract Infection ◽

Antimicrobial Agents ◽

Hospitalized Patients ◽

Diffusion Method ◽

Disk Diffusion Method ◽

E Coli

Background and Objectives: Urinary tract infection (UTI) is one of the most frequent infectious diseases which is caused by Gram-negative bacteria especially Escherichia coli. Multiple resistance to antimicrobial agents are increasing quickly in E. coli isolates and may complicate therapeutic strategies for UTI. The propose of this study was to determine the antibiotic resistance patterns and the multidrug-resistance (MDR) phenotypes in uropathogenic E. coli (UPEC). Materials and Methods: A total of 153 UPEC isolates were collected from both hospitalized patients (95 isolates) and outpatients (58 isolates) from March to October 2018. In order to determine the MDR among UPEC isolates, we have tested 15 antimicrobial agents on Muller Hinton agar by the disk diffusion method. Results: The percentage of MDR isolates (resistant to at least three drug classes such as fluoroquinolones, penicillins and cephalosporins) was 55.5% in the hospitalized patients and the outpatients. Antibiotic resistance to ampicillin, ceftazidime, nalidixic acid and trimethoprim/ sulfamethoxazole was higher than 60%. Meropenem, Imipenem and norfloxacin indicated markedly greater activity (93.3%, 80% and 85.6%, respectively) than other antimicrobial agents. Conclusions: Urinary tract infection due to MDR E. coli may be difficult to treat empirically due to high resistance to commonly used antibiotics, so, empirical antibiotic treatment should be reviewed periodically at local studies.

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Prevalence and antibiotic resistance profile of Shiga toxin-producing Escherichia coli isolated from diarrheal samples

Iranian Journal of Microbiology ◽

10.18502/ijm.v12i4.3931 ◽

2020 ◽

Author(s):

Farzad Esavand Heydari ◽

Mojtaba Bonyadian ◽

Hamdallah Moshtaghi ◽

Masoud Sami

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Shiga Toxin ◽

Virulence Genes ◽

Enterohemorrhagic Escherichia Coli ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Resistance Profile ◽

E Coli ◽

Antibiotic Resistance Profile

Background and Objectives: Enterohemorrhagic Escherichia coli (EHEC) causes bloody and non-bloody diarrhea, intestinal infection and extraintestinal complications in humans. This study aimed to isolate and evaluate the prevalence of E. coli O157: H7 and other Shiga toxin-producing E. coli (STEC) and identify the virulence genes (stx1, stx2, hly and eaeA) from patients with diarrhea. Also, the antibiotic resistance profile of the isolated strains was evaluated. Materials and Methods: A total of 100 stool samples were collected from patients with acute diarrhea referring to the hospital and clinics in Isfahan County, Iran. Phenotypic tests and PCR assay were used for detection of E. coli O157: H7 and other Shiga toxin-producing E. coli. The presence of virulence genes (stx1, stx2, hly and eaeA) were identified by PCR. The antibiotic resistance profile of the isolates was determined using the agar disk diffusion method. The results were analyzed descriptively by Sigma stat version 4 software. Results: Seventy - eight out of 100 samples (78%) were contaminated with E. coli. E. coli O157 was isolated from five samples (6.4%), of which only two strains (2.56%) were identified as E. coli O157: H7. According to the results, out of two E. coli O157: H7 isolates, one (50%) isolate contained eaeA and two isolates (100%) contained Stx1, Stx2, hlyA genes. Out of three (3.84%) E. coli O157: HN, one of the isolate (33.3%) contained stx1 and, two isolates (66.7%) were positive for hlyA genes. Also, the results revealed that six strains (7.69%) were non-O157: H7 STEC, of which two isolates (33.3%) contained stx1 and four isolates (66.7%) were positive for stx2 and hlyA genes. The results of antibiogram tests revealed that all of the STEC isolates (100%) were sensitive to imipenem followed by kanamycin, gentamicin and nitrofurantoin (91%). High resistance (54.5%) to ampicillin and ciprofloxacin was observed among the STEC isolates. Conclusion: The results of the current study showed that although the prevalence of E. coli O157: H7 was low among patients with diarrhea, the other STEC strains with relative resistance to antibiotics are more prevalent.

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