Gene Sequencing of Gelatinase among E. faecalis Isolates
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Keywords
PCR, Enterococcus faecalis, gelE gene, UTIs.
Abstract
Background: Enterococcus is a genus of Gram-positive, catalase-negative, non-spore-forming, facultatively anaerobic bacteria that may exist alone or in chains. The lactic acid bacteria (LAB) that generate bacteriocins include the enterococci. gelE on the E. faecalis chromosome codes for gelatinase, a zinc-containing metalloproteinase that is released.
Objective: Virulence gene sequencing for gelatinase in E. faecalis strains
Materials and methods: Patients hospitalized and seen at Baghdad's Al-Karama Hospital and Medical City Hospital throughout a three-month period (May to July 2022) provided the study's 200 participants with urine and vaginal specimens. The samples were cultured for (18-24) hours in various mediums. Then, they were incubated at (37oC) for (18-24) hours on a number of different selective media. When first trying to identify E. faecalis, scientists relied on colony morphology, microscopic examinations, and biochemical assays.
Results: All 200 clinical samples cultured positive, however only 44(22%) of the isolates were associated with E. faecalis, the automated Vitek 2 system employed GP-ID cards containing 64 biochemical assays to ensure the isolates were really E. faecalis. Using this method, it was able to quickly identify 44 different bacterial isolates, with a confidence level ranging from excellent (probability percentage of (94 to 99.7%).The virulence factor gelatinase gene in E. faecalis was studied in 47 isolates of the bacteria obtained from various environments. Results from our analysis indicated that, only 14 of the 44 isolates (31.8%) tested positive for this gene, with a molecular length of 213bp. The sequencing of gelE gene shows for E. faecalis having one transversion A/C, and the effect Missense. From the Gene Bank, found part of gelE gene having 99% compatibility with the subject of gelE gene in NCBI under sequence ID: CP0881981. Another part of sequencing for gelE gene to E. faecalis, the results shows compatibility of 100% in Gene Bank of gelE under sequence ID: CP088198.1, so no recorded change noticed from the gene in this isolate. Furthermore, neighbour phylogenetic distances in this tree indicated a wide biological diversity of E. faecalis sequences.
Conclusions: The pathogenicity of E. faecalis increased by presence of gelE gene.
References
2. Deng J J, Deng D, Wang Z L, Luo X C, Chen H P, Liu S Y & Li J Z. Indole Metabolism Mechanisms in a New, Efficient Indole-degrading Facultative Anaerobe Isolate Enterococcus hirae GDIAS-5. Journal of Hazardous Materials, 2022; 128890.
3. Glennon-Alty L, Hackett A P, Chapman E A & Wright H L. Neutrophils and redox stress in the pathogenesis of autoimmune disease. Free Radical Biology and Medicine, 2018; 125, 25-35.
4. Yong A M H. HtrA and CroRS two-component signal transduction system monitor sortase-assembled pilus biogenesis in Enterococcus faecalis (Doctoral dissertation, Nanyang Technological University). 2019.
5. Singh K V, Pinkston K L, Gao P, Harvey B R & Murray B E. Anti-Ace monoclonal antibody reduces Enterococcus faecalis aortic valve infection in a rat infective endocarditis model. Pathogens and Disease, 2018; 76(8), fty084.
6. Yousuf, B, Adachi K & Nakayama J. Developing Anti-virulence Chemotherapies by Exploiting the Diversity of Microbial Quorum Sensing Systems. In Biotechnological Applications of Quorum Sensing Inhibitors 2018; 151-208. Springer, Singapore.
7. He Z, Liang J, Zhou W, Xie Q, Tang Z, Ma R & Huang Z.. Effect of the quorum‐sensing luxS gene on biofilm formation by Enterococcus faecalis. European Journal of Oral Sciences, 2016; 124(3), 234-240.
8. Ali, L., Goraya, M. U., Arafat, Y., Ajmal, M., Chen, J. L., & Yu, D. (2017). Molecular mechanism of quorum-sensing in Enterococcus faecalis: its role in virulence and therapeutic approaches. International journal of molecular sciences, 18(5), 960.
9. Ali L, Mustafa M, Xiao Z R, Islam W, Ara U, Ajmal M & Yu D. Responses of Enterococcus faecalis resistance and cytolysin up-regulation to nutrients in constructed mesocosms. Journal of King Saud University-Science, 2022; 34(1), 101680.
10. Ames B N. Dietary carcinogens and anticarcinogens: oxygen radicals and degenerative diseases. Science, 1983; 221(4617), 1256-1264.
11. Aghdam M A, Barhaghi M S, Aghazadeh M, Jafari F, Hagh M B, Haghdoost M & Kafil H S. Virulence genes in biofilm producer Enterococcus faecalis isolates from root canal infections. Cellular and Molecular Biology, 2017; 63(5), 55-59.
12. Hashem Y A, Abdelrahman K A & Aziz R K. Phenotype–Genotype Correlations and Distribution of Key Virulence Factors in Enterococcus faecalis Isolated from Patients with Urinary Tract Infections. Infection and Drug Resistance, 2021; 14, 1713.
13. Stępień-Pyśniak D, Bertelloni F, Dec M, Cagnoli G, Pietras-Ożga D, Urban-Chmiel R & Ebani V V. Characterization and Comparison of Enterococcus spp. Isolates from Feces of Healthy Dogs and Urine of Dogs with UTIs. Animals, 2021; 11(10), 2845.
14. Jahansepas A, Sharifi Y, Aghazadeh M & Ahangarzadeh Rezaee M. Comparative analysis of Enterococcus faecalis and Enterococcus faecium strains isolated from clinical samples and traditional cheese types in the Northwest of Iran: Antimicrobial susceptibility and virulence traits. Archives of microbiology, 2020; 202(4), 765-772.
15. Wójkowska-Mach J, Pomorska-Wesołowska M, Romanik M & Romaniszyn D. Prevalence and antimicrobial susceptibility profiles of microorganisms associated with lower reproductive tract infections in women from southern poland—Retrospective laboratory-based study. International Journal of Environmental Research and Public Health, 2021; 18(1), 335.
16. Mancuso G, Midiri A, Gerace E & Biondo C. Bacterial antibiotic resistance: the most critical pathogens. Pathogens, 2021; 10(10), 1310.
17. Giannakopoulos X, Sakkas H, Ragos V, Tsiambas E, Bozidis P, Evangelou A M & Sofikitis N. Impact of enterococcal urinary tract infections in immunocompromised-neoplastic patients. J BUON, 2019; 24(5), 1768-1775.
18. Majumder M M I, Ahmed T, Ahmed S & Khan A R. Microbiology of catheter associated urinary tract infection. In Microbiology of Urinary Tract Infections-Microbial Agents and Predisposing Factors. IntechOpen. 2018.
19. Toc D A, Pandrea, S L, Botan A, Mihaila R M, Costache C A, Colosi I A & Junie L M. Enterococcus raffinosus, Enterococcus durans and Enterococcus avium Isolated from a Tertiary Care Hospital in Romania—Retrospective Study and Brief Review. Biology, 2022; 11(4), 598.
20. Ahmed S H & Hafidh R R. The Isolation of specifically lytic phages along with their extracted endolysins as antibacterial agents to MDR Enterococcus faecalis. Research Journal of Pharmacy and Technology, 2021; 14(9), 4547-4554.
21. da Silva R A, Tay W H, Ho F K, Tanoto F R, Chong K K, Choo P Y & Kline K A. Enterococcus faecalis alters endo-lysosomal trafficking to replicate and persist within mammalian cells. PLoS pathogens, 2022; 18(4), e1010434.
22. Weiner-Lastinger L M, Abner S, Edwards J R, Kallen A J, Karlsson M, Magill S S & Dudeck M A. Antimicrobial-resistant pathogens associated with adult healthcare-associated infections: summary of data reported to the National Healthcare Safety Network, 2015–2017. Infection Control & Hospital Epidemiology, 2020; 41(1), 1-18.
23. Wang Y, Liang Q, Lu B, Shen H, Liu S, Shi Y & Chen H. Whole-genome analysis of probiotic product isolates reveals the presence of genes related to antimicrobial resistance, virulence factors, and toxic metabolites, posing potential health risks. BMC genomics, 2021; 22(1), 1-12.
24. Farman M, Yasir M, Al-Hindi R R, Farraj S A, Jiman-Fatani A A, Alawi M & Azhar E I. Genomic analysis of multidrug-resistant clinical Enterococcus faecalis isolates for antimicrobial resistance genes and virulence factors from the western region of Saudi Arabia. Antimicrobial Resistance & Infection Control, 2019; 8(1), 1-11.
25. Esmail M A M, Abdulghany H M & Khairy R M. Prevalence of multidrug-resistant Enterococcus faecalis in hospital-acquired surgical wound infections and bacteremia: Concomitant analysis of antimicrobial resistance genes. InfectiousDiseases: Research and Treatment, 2019; 12, 1178633719882929.
26. Kaviar V H, Khoshnood S, Asadollahi P, Kalani B S, Maleki A, Yarahmadi S & Pakzad I. Survey on phenotypic resistance in Enterococcus faecalis: comparison between the expression of biofilm-associated genes in Enterococcus faecalis persister and non-persister cells. Molecular biology reports, 2022; 49(2), 971-979.
27. Iseppi R, Di Cerbo, A, Messi P & Sabia C. Antibiotic resistance and virulence traits in vancomycin-resistant enterococci (Vre) and extended-spectrum β-lactamase/ampc-producing (ESBL/ampc) enterobacteriaceae from humans and pets. Antibiotics, 2020; 9(4), 152.
28. Olvera-Rosales L B, Cruz-Guerrero A E, García-Garibay J M, Gómez-Ruíz L C, Contreras-López E, Guzmán-Rodríguez F & González-Olivares L G. Bioactive peptides of whey: obtaining, activity, mechanism of action, and further applications. Critical Reviews in Food Science and Nutrition, 2022; 1-31.
29. Gopalasamy K & Geetha R V. Genotypic characterization of Enterococcus faecalis isolated from patient undergoing endodontic treatment. Drug Invention Today, 2018; 10.
30. Liesenborghs L, Meyers S, Vanassche T & Verhamme P. Coagulation: At the heart of infective endocarditis. Journal of Thrombosis and Haemostasis, 2020; 18(5), 995-1008.
31. Willett J L, Robertson E B & Dunny G M. The phosphatase Bph and peptidyl-prolyl isomerase PrsA are required for gelatinase expression and activity in Enterococcus faecalis. Journal of Bacteriology, 2022; e00129-22.
32. Ferchichi M, Sebei K, Boukerb A M, Karray-Bouraoui N, Chevalier S, Feuilloley, M G & Zommiti M. Enterococcus spp.: Is It a Bad Choice for a Good Use—A Conundrum to Solve?. Microorganisms, 2021; 9(11), 2222.
33. Sneath P.H.A. and Sokal R.R. Numerical Taxonomy. Freeman, San Francisco. 1973
34. Tamura K, Nei M, and Kumar S. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proceedings of the National Academy of Sciences (USA) 2004; 101:11030-11035.
35. Tamura K, Stecher G, Peterson D, Filipski A and Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution30: 2725-2729. 2013.
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