Molecular Identification of Biofilm-forming Pseudomonas aeruginosa Isolates from Noncritical Surfaces of a Tertiary Healthcare Center in Abia State, Southeast Nigeria

Authors

  • Ejiofor C. Agbo Department of Medical Laboratory Services, Federal Medical Centre, Umu Obasi 440236, Umuahia, Nigeria.
  • Obiora. R. Ejinaka Department Parasitology, Federal School of Medical Laboratory Science, Jos university Teaching Hospital Nigeria, Jos, Nigeria.
  • Obiorah E. Okechukwu Department of Medical Laboratory Services, Federal Medical Centre, Umu Obasi 440236, Umuahia, Nigeria.
  • Obeta M. Uchejeso Department of Medical Laboratory Management, Federal School of Medical Laboratory Science, Jos university Teaching Hospital Nigeria, Jos, Nigeria.
  • Jwanse I. Rinpan Health and Development Support Programme (HANDS), Jos, Nigeria.
  • Ifeoma E Lote-Nwaru Department of Medical Microbiology, Federal School of Medical Laboratory Science, Jos university Teaching Hospital Nigeria, Jos, Nigeria.

DOI:

https://doi.org/10.54987/jobimb.v10i1.659

Keywords:

Molecular characterization, Healthcare-associated infections, Biofilm forming, Pseudomonas species, Tertiary Medical Centre

Abstract

The presence of biofilm-forming Pseudomonas aeruginosa on noncritical surfaces in a hospital has been attributed to healthcare-associated infections (HAIs) leading to multidrug-resistant (MDR) infections. This study is aimed at identifying the molecular characteristics of Biofilm forming Pseudomonas aeruginosa found on non-critical surfaces in a hospital environment. Samples were collected using the swab technique from different noncritical surfaces surrounding hospitalized patients in different wards namely sphygmomanometers, thermometers, bed rails etc. Bacterial analysis was performed by using conventional microbiological techniques, biochemical tests and Microbact 24E assay. Biofilm forming Pseudomonas aeruginosa were visualized using Crystal Violet staining assay and then estimated by measuring the Optical Density through Spectrophotometer. Four hundred and fifty (450) positive samples of bacterial isolates from noncritical surfaces of six selected wards of the hospital were obtained from one thousand three hundred and fourteen (1314) samples. One hundred and thirty-nine (139) isolates of Pseudomonas spp. were obtained, out of which 115 (82.73%) were identified as biofilm formers. Of these 115 biofilm formers, 83(72.17%) belonged to Pseudomonas aeruginosa, 30(22.09) Pseudomonas fluorescens, 20(17.39%) Pseudomonas putida and 6(5.22%) belongs to Pseudomonas chlorophis. The final characterization of Biofilm forming Pseudomonas aeruginosa isolates was verified by 16S rDNA sequencing. Sequence analysis showed similarity at 99.5% Pearson`s correlation coefficient (r) between 16S rDNA of the representative standard isolate (from gene library) and Biofilm forming Pseudomonas aeruginosa strain obtained from the Tertiary health facility, Federal Medical Centre (FMC), Umuahia. The nucleotide sequence of the experimental isolates expressed genetic mutation which represents the point of involvement of Pel B gene and is involved in the biofilm formation character of Pseudomonas aeruginosa in this study. There is an urgent need to take nosocomial transmission from noncritical surfaces serious in FMC Umuahia.

References

Festini F, Buzzetti R, Bassi C, Braggion C, Salvatore D, Taccetti G, et al. Isolation measures for prevention of infection with respiratory pathogens in cystic fibrosis: a systematic review.

Araújo P, Lemos M, Mergulhão F, Melo L, Simões M. Antimicrobial resistance to

Bell M. Biofilms: A Clinical Perspective. Curr Infect Dis Rep. 2001;3(6):483-6.

Karatan E, Watnick P. Signals, regulatory networks, and materials that build and break

Nseir S, Blazejewski C, Lubret R, Wallet F, Courcol R, Durocher A. Risk of acquiring

Ryder C, Byrd M, Wozniak DJ. Role of polysaccharides in Pseudomonas aeruginosa biofilm development. Curr Opin Microbiol. 2007;10(6):644-8.

Ma B, Chen H, He Y, Wang H, Xu J. Evaluation of toxicity risk of polycyclic aromatic

Sutherland IW. The biofilm matrix--an immobilized but dynamic microbial environment. Trends Microbiol. 2001;9(5):222-7.

Franklin M, Nivens D, Weadge J, Howell P. Biosynthesis of the Pseudomonas aeruginosa

Byrd MS, Sadovskaya I, Vinogradov E, Lu H, Sprinkle AB, Richardson SH, et al. Genetic and biochemical analyses of the Pseudomonas aeruginosa Psl exopolysaccharide reveal overlapping roles for polysaccharide synthesis enzymes in Psl and LPS production. Mol

Swartjes JJTM, Das T, Sharifi S, Subbiahdoss G, Sharma PK, Krom BP, et al. A functional dnase i coating to prevent adhesion of bacteria and the formation of biofilm. Adv Funct

Yang L, Barken KB, Skindersoe ME, Christensen AB, Givskov M, Tolker-Nielsen T. Effects

Gloag ES, Turnbull L, Huang A, Vallotton P, Wang H, Nolan LM, et al. Self-organization

Allesen-Holm M, Barken KB, Yang L, Klausen M, Webb JS, Kjelleberg S, et al. A

Finkel SE, Kolter R. Dna as a nutrient: novel role for bacterial competence gene homologs. J Bacteriol. 2001;183(21):6288-93.

O'Toole GA, Kolter R. Flagellar and twitching motility are necessary for Pseudomonas

Stepanovi? S, Cirkovi? I, Ranin L, Svabi?-Vlahovi? M. Biofilm formation by Salmonella

Collin C, Lyne P. Collins and Lyne's Microbiological methods. 8th ed. London: Arnold

Mailafia S, Olaaatunde H, Godspower O, Jacobs C, Shuaibu G, Onyilokwu S. Microbact

Shukla SK, Rao TS. An Improved Crystal Violet Assay for Biofilm Quantification in 96-

Spilker T, Coenye T, Vandamme P, LiPuma JJ. Pcr-based assay for differentiation of

Frank JA, Reich CI, Sharma S, Weisbaum JS, Wilson BA, Olsen GJ. Critical evaluation of

O'Toole GA. Microtiter Dish Biofilm Formation Assay. J Vis Exp JoVE. 2011;(47):2437.

Hernández J, Ferrus MA, Hernández M, Owen RJ. Arbitrary primed PCR fingerprinting and

Pontes DS, Pinheiro FA, Lima-Bittencourt CI, Guedes RLM, Cursino L, Barbosa F, et al. Multiple antimicrobial resistance of gram-negative bacteria from natural oligotrophic lakes

Sebastian A, Larsson L. Characterization of the microbial community in indoor

Friedman L, Kolter R. Genes involved in matrix formation in Pseudomonas aeruginosa

Rutala WA and David J. Weber. Guideline for disinfection and sterilization in healthcare facilities, Hospital Epidermiology, 2008; NC 27514.

Downloads

Published

31.07.2022 — Updated on 02.08.2022

Versions

How to Cite

Agbo, . E. C., Ejinaka, O. R., Okechukwu, O. E. ., Uchejeso, O. M. ., Rinpan, J. I. ., & Lote-Nwaru, . I. E. (2022). Molecular Identification of Biofilm-forming Pseudomonas aeruginosa Isolates from Noncritical Surfaces of a Tertiary Healthcare Center in Abia State, Southeast Nigeria. Journal of Biochemistry, Microbiology and Biotechnology, 10(1), 25–30. https://doi.org/10.54987/jobimb.v10i1.659 (Original work published July 31, 2022)

Issue

Vol. 10 No. 1 (2022)

Section

Articles

License

Copyright (c) 2022 Journal of Biochemistry, Microbiology and Biotechnology

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Authors who publish with this journal agree to the following terms:

    1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
    2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
    3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).