Biofilm formation, antimicrobial susceptibility and virulence genes of Uropathogenic Escherichia coli isolated from clinical isolates in Uganda

Background: Uropathogenic Escherichia coli (UPEC) strains, encoding superficial and secretory virulence factors, can lead to colonization and facilitation of bacterial growth in the host urinary tract, causing Urinary Tract Infection (UTI). Objectives: This study determined the ability of biofilm formation by the Congo red agar (CRA) method, the presence of virulence genes using the multiplex polymerase chain reaction (PCR) method, and the relationship between biofilm formation and antibiotic resistance patterns and virulence genes in E. coli clinical isolates in Yasuj. Methods: This cross-sectional study was performed on 144 UPEC isolates collected in 2017. Biofilm formation was detected by the CRA phenotypic assay and virulence factors by the multiplex PCR method. Antibiotic resistance tests were performed by the Kirby-Bauer method. Results: Out of 144 isolates of E. coli, 22 (19.4%) isolates showed to be strong biofilm producers, 27 (23.8%) moderate biofilm producers, and 64 (56.3%) weak biofilm producers. A significant relationship was observed between biofilm-producing strains and resistance to ampicillin (P = 0.020) and cotrimoxazole (P = 0.038). The virulence genes in strong biofilm producers included iutA (95%), FimH (93%), ompT (90%), PAI (90%), and TraT (81%) genes. The phylogroup B2 carried the most virulence genes. A significant correlation was observed between E. coli phylogenetic groups and aer (P = 0.019), iroN (P = 0.042), and ompT (P = 0.032) virulence genes. Conclusions: The results of this study showed a high prevalence of virulence genes, and antibiotic-resistant E. coli strains capable of biofilm formation. The results of this study may help elucidate the pathogenesis of UPEC and facilitate better treatment strategies for patients with UTIs in this geographic area.

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Draft Genome Sequences of Semiconstitutive Red, Dry, and Rough Biofilm-Forming Commensal and Uropathogenic Escherichia coli Isolates

Genome Announcements ◽

10.1128/genomea.01249-16 ◽

2017 ◽

Vol 5 (4) ◽

Cited By ~ 3

Author(s):

Annika Cimdins ◽

Petra Lüthje ◽

Fengyang Li ◽

Irfan Ahmad ◽

Annelie Brauner ◽

...

Keyword(s):

Escherichia Coli ◽

Biofilm Formation ◽

Molecular Basis ◽

Draft Genome ◽

Uropathogenic Escherichia Coli ◽

Clinical Isolates ◽

Genome Sequences ◽

Content Type ◽

E Coli ◽

K 12

ABSTRACT Strains of Escherichia coli exhibit diverse biofilm formation capabilities. E. coli K-12 expresses the red, dry, and rough (rdar) morphotype below 30°C, whereas clinical isolates frequently display the rdar morphotype semiconstitutively. We sequenced the genomes of eight E. coli strains to subsequently investigate the molecular basis of semiconstitutive rdar morphotype expression.

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Biofilm formation, antimicrobial susceptibility and virulence genes of Uropathogenic Escherichia coli isolated from clinical isolates in Uganda

10.21203/rs.2.19141/v2 ◽

2020 ◽

Author(s):

Paul Katongole ◽

Fatuma Nalubega ◽

Najjuka Christine Florence ◽

Benon Asiimwe ◽

Irene Andia

Keyword(s):

Antimicrobial Resistance ◽

Antimicrobial Susceptibility ◽

Biofilm Formation ◽

Virulence Genes ◽

Clinical Isolates ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Cross Sectional ◽

E Coli ◽

Tract Infections

Abstract Introduction: Uropathogenic E. coli is the leading cause of Urinary tract infections (UTIs), contributing to 80-90% of all community-acquired and 30-50% of all hospital-acquired UTIs. Biofilm forming Uropathogenic E. coli are associated with persistent and chronic inflammation leading to complicated and or recurrent UTIs. Biofilms provide an environment for poor antibiotic penetration and horizontal transfer of virulence genes which favors the development of Multidrug-resistant organisms (MDRO). Understanding biofilm formation and antimicrobial resistance determinants of Uropathogenic E. coli strains will provide insight into the development of treatment options for biofilm-associated UTIs. The aim of this study was to determine the biofilm forming capability, presence of virulence genes and antimicrobial susceptibility pattern of Uropathogenic E. coli isolates in Uganda. Methods: This was a cross-sectional study carried in the Clinical Microbiology and Molecular biology laboratories at the Department of Medical Microbiology, Makerere University College of Health Sciences. We randomly selected 200 Uropathogenic E. coli clinical isolates among the stored isolates collected between January 2018 and December 2018 that had significant bacteriuria (>105 CFU). All isolates were subjected to biofilm detection using the Congo Red Agar method and Antimicrobial susceptibility testing was performed using the Kirby disk diffusion method. The isolates were later subjected PCR for the detection of Urovirulence genes namely; Pap, Fim, Sfa, Afa, Hly and Cnf, using commercially designed primers.Results: In this study, 62.5% (125/200) were positive biofilm formers and 78% (156/200) of these were multi-drug resistant (MDR). The isolates were most resistant to Trimethoprim sulphamethoxazole and Amoxicillin (93%) followed by gentamycin (87%) and the least was imipenem (0.5%). Fim was the most prevalent Urovirulence gene (53.5%) followed by Pap (21%), Sfa (13%), Afa (8%), Cnf (5.5%) and Hyl (0%).Conclusions: We demonstrate a high prevalence of biofilm-forming Uropathogenic E. coli strains that are highly associated with the MDR phenotype. We recommend routine surveillance of antimicrobial resistance and biofilm formation to understand the antibiotics suitable in the management of biofilm-associated UTIs.

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Biofilm formation, antimicrobial susceptibility and virulence genes of Uropathogenic Escherichia coli isolated from clinical isolates in Uganda

10.21203/rs.2.19141/v1 ◽

2019 ◽

Author(s):

Paul Katongole ◽

Fatuma Nalubega ◽

Najjuka Christine Florence ◽

Benon Asiimwe ◽

Irene Andia

Keyword(s):

Antimicrobial Resistance ◽

Antimicrobial Susceptibility ◽

Biofilm Formation ◽

Virulence Genes ◽

Clinical Isolates ◽

Diffusion Method ◽

Disk Diffusion Method ◽

Cross Sectional ◽

E Coli ◽

Tract Infections

Abstract Introduction: Uropathogenic E. coli is the leading cause of Urinary tract infections (UTIs), contributing to 80-90% of all community-acquired and 30-50% of all hospital-acquired UTIs. Biofilm forming Uropathogenic E. coli are associated with persistent and chronic inflammation leading to complicated and or recurrent UTIs. Biofilms provide an environment for poor antibiotic penetration and horizontal transfer of virulence genes which favors the development of Multidrug-resistant organisms (MDRO). Understanding biofilm formation and antimicrobial resistance determinants of Uropathogenic E. coli strains will provide insight into the development of treatment options for biofilm-associated UTIs. The aim of this study was to determine the prevalence of biofilm formation among Uropathogenic E. coli clinical isolates, their relationship with antimicrobial susceptibility patterns, and Urovirulence genes. Methods: This was a cross-sectional study carried in the Clinical Microbiology and Molecular biology laboratories at the Department of Medical Microbiology, Makerere University College of Health Sciences. We randomly selected 200 Uropathogenic E. coli clinical isolates among the stored isolates collected between January 2018 and December 2018 that had significant bacteriuria (>105 CFU). All isolates were subjected to biofilm detection using the Congo Red Agar method and Antimicrobial susceptibility testing was performed using the Kirby disk diffusion method. The isolates were later subjected PCR for the detection of Urovirulence genes namely; Pap, Fim, Sfa, Afa, Hly and Cnf, using commercially designed primers.Results: In this study, 62.5% (125/200) were positive biofilm formers and 78% (156/200) of these were multi-drug resistant(MDR). The isolates were most resistant to Trimethoprim sulphamethoxazole and Amoxicillin (93%) followed by gentamycin (87%) and the least was imipenem (0.5%). Fim was the most prevalent Urovirulence gene (53.5%) followed by Pap (21%), Sfa (13%), Afa (8%), Cnf (5.5%) and Hyl (0%).Conclusions: We demonstrate a high prevalence of biofilm-forming Uropathogenic E. coli strains that are highly associated with the MDR phenotype. We recommend routine surveillance of antimicrobial resistance and biofilm formation to understand the antibiotics suitable in the management of biofilm-associated UTIs.

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Molecular Analysis of Uropathogenic E.coli Isolates from Urinary Tract Infections

Infectious Disorders - Drug Targets ◽

10.2174/1871526518666181113091322 ◽

2019 ◽

Vol 19 (3) ◽

pp. 322-326 ◽

Cited By ~ 1

Author(s):

Hassan Valadbeigi ◽

Elham Esmaeeli ◽

Sobhan Ghafourian ◽

Abbas Maleki ◽

Nourkhoda Sadeghifard

Keyword(s):

Escherichia Coli ◽

Polymerase Chain Reaction ◽

Urinary Tract ◽

Urinary Tract Infections ◽

Multiplex Polymerase Chain Reaction ◽

Virulence Genes ◽

Uropathogenic Escherichia Coli ◽

Chain Reaction ◽

Polymerase Chain ◽

Tract Infections

Introduction: The aim of the current study was to investigate the prevalence of virulence genes in uropathogenic Escherichia coli (UPEC) isolates in Ilam. Materials and Methods: For this purpose, a total of 80 UPEC isolates were collected for patients with UTIs during a 6 months period. The multiplex polymerase chain reaction (multiplex PCR) was used to detect the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. Results: The prevalence of fimH, papEF, iucD, fyuA, hlyA, hlyA, and ompT genes were 87.5%, 47.5%, 60%, 67.5%, 27.5%, 47.5% and 71.2%, respectively. Among all of the isolates, 27 profiles were obtained. Conclusion: Our findings demonstrated that the most prevalence was found for fimH, and different distribution of virulence genes suggested different ability of pathogenicity.

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