Detecting of Biofilm Formation and Associated Genes, and Assessing PilA Gene Expression in Different Pseudomonas aeruginosa Wounds and Burns Isolated in Baghdad Hospitals
Keywords:
Biofilm, Biofilm, Pseudomonas aeruginosa, Pseudomonas aeruginosa, genes pil A, genes pil A, Psl A, Psl A, Psl B, Psl B, RHII and rhIAB, RHII and rhIAB, gene expression, gene expressionAbstract
A total of 103 clinical specimens were obtained from patients have burns, and wounds at ages between 10 - 60 years from both genders in three hospitals in Baghdad – Iraq: Al Yarmouk Teaching Hospital, Baghdad Teaching Hospital, and Al-Karkh General Hospital and at the period from (23/9/2022 to 29/12/2022). Pseudomonas aeruginosa was isolated and identified, which showed that from a total of 130 specimens, only 103 isolates (71.87%) belonged to P. aeruginosa. The isolation rate was higher from burn infections followed by wound infections, at ratios 40.62%, and 23.75%, respectively, the rest 27 isolates (16.87%) were related to other bacterial spp. like Escherichia coli, Klebsiella pneumonia, Proteus mirabilis, Staphylococcus aureus and Enterobacter cloacae. The results showed that all P. aeruginosa isolates were a multidrug resistance to many antibiotics. Estimating the growth for isolates at different time (6, 12, 18, 24, and 36) hrs. revealed that a high OD value was detected between 18-24 hrs.
The results of the molecular study to detect biofilm associated genes pil A, Psl A, Psl B, RHII and rhIAB revealed that these genes were detected in P. aeruginosa isolates at ratio (93%, 80%, 90%, 90%, and 86.6%) respectively. Also, the detection of gene expression for PilA gene in three P. aeruginosa isolates at different time 6, 12, 18, 24, and 36 hrs. by RT-PCR revealed that a high gene expression of pilA gene in P. aeruginosa isolates detected between 18-24 hrs.
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Abbasi A., Maddah S. M., Mahboubi A., Khaledi A., Vazini H., Esmaeili D. (2017). Investigate the inhibitory effects of Satureja khuzestanica essential oil against housekeeping fabD and exoA genes of Pseudomonas aeruginosa from hospital isolates using RT-PCR technique. Annals of Medical and Health Sciences Research, 7(4), 246-250.
Abd El Galil, K., AbdelGhani, S. M., Sebak, M. A. and El-Naggar, W., (2013). Detection of biofilm genes among clinical isolates of Pseudomonas aeruginosa recovered from some Egyptian hospitals. Journal. Microbiology, 36, 86-101.
Abdelraheem, W. M.; and Mohamed, E. S. (2021). The effect of zinc oxide nanoparticles on Pseudomonas aeruginosa biofilm formation and virulence genes expression. The Journal of Infection in Developing Countries, 15(06), 826-832Abdelraheem W. M., Abdelkader A. E., Mohamed E., Mohammed M. (2020). Detection of biofilm formation and assessment of genes expression in different Pseudomonas aeruginosa clinical isolates. Meta Gene, 23, 100646.
Algammal A. M., Mabrok M., Ramasamy E., Youssef F., Atwa M., El‑kholy A., Hetta H., Hozzein W. (2020). Zoology Department, College of Science, King Saud University, Riyadh, Saudi Arabia. Botany and Microbiology Department, Faculty of Science, Beni-Suef University, Beni‑Suef 62511, Egypt.
Al-Mayyahi A. W. (2018). Detection of (exoT, exoY, exo S and exoU) Genes in Pseudomonas aeruginosa isolate from different clinical sources (MSc.), Institute of Genetic Engineering and Biotechnology for Post Graduate Studies, University of Baghdad. 60-63.
Breijyeh, Z.; Jubeh, B. and Karaman, R. (2020). Resistance of Gram-negative bacteria to current antibacterial agents and approaches to resolve it. Molecul., 25 (1340): 1-23. doi:10.3390molecules25061340.
Bilici K., Atac N., Muti A., Baylam I., Dogan O., Sennaroglu A., Can F., Acar H. (2020). Broad spectrum antibacterial photodynamic and photothermal therapy achieved with indocyanine green loaded SPION sunder near infrared irradiation. Biomater. Sci, 8, 4616–4625.biofilm and its role in the pathogenesis of disease. Antibiotics, 9(2), 59
Brandenburg K. S., Weaver A. J., Karna S., You T., Chen P., Stryk S., Leung K. (2019). Formation of Pseudomonas aeruginosa biofilms in full-thickness scald burn wounds in rats. Scientific reports, 9(1), 1-12.
Breijyeh et al., 2020
Cappuccino J. G., Welsh C. (2018). Microbiology A laboratory manual. 11th ed. Pearson Education Limited Edinburgh Gate Harlow Essex CM20 2JE. England.
Caschera B. W., Dassi C., Vivekanandan A., Zoing M., Douglas R. G., Casilag F., Lorenz A., et al. (2016). The LasB elastase of Pseudomonas aeruginosa acts in concert with alkaline protease AprA to prevent flagellin-mediated immune recognition. Infection and immunity, 84(1): 162-171.
Cigana C., Castandet J., Sprynski N., Melessike M., Beyria L., Ranucci S., Everett M. (2021). Pseudomonas aeruginosa elastase contributes to the establishment of chronic lung colonization and modulates the immune response in a murine model. Frontiers in microbiology, 11, 620819.
Corley J. M., Intile P., Yahr T. (2022). Direct Inhibition of RetS Synthesis by RsmA Contributes to Homeostasis of the Pseudomonas aeruginosa Gac/Rsm Signaling System. Journal of Bacteriology, 204(3), e00580-21.
Correia S., Poeta P., Hébraud M., Capelo J., Igrejas G. (2017). Mechanisms of quinolone action and resistance: where do we stand? Journal of Medical Microbiology, 66(5), 551-559.
Costa O. Y., Raaijmakers J. M., Kuramae E. (2018). Microbial extracellular polymeric substances: ecological function and impact on soil aggregation. Front. Microbiol. 9, 1636.
Challapilli, S.B. (2018). Recent perspectives on the molecular basis of biofilm formation by Pseudomonas aeruginosa and approaches for treatment and biofilm dispersal. Folia. Microbiol. 63(4):413-432.
Divyashree, M., Mani, M. K., Shama, P. K., Vijaya, K. D., Veena, S. A., Shetty, A. K. & Karunasagar, I. 2020 Hospital wastewater treatment reduces NDM-positive bacteria being discharged into water bodies. Water Environment Research 92 (4), 562–568.
Dai M., Wang S., Wang T., Hu S., Bhayana B., Ishii M., Kong Y., Cai Y., Lu T., Cui W. (2021). Bacteria-specific phototoxic reactions triggered by blue light and phytochemical carvacrol. Sci. Transl. Med. 2021, 13, 575.
Di Martino P. (2018). Extracellular Polymeric Substances, a Key Element in Understanding Biofilm Phenotype. AIMS Microbiol. 4, 274–288.
Durant D. J. (2017). Nurse-driven protocols and the prevention of catheter- associated urinary tract infections: A systematic review. Am J Infect Control;45(12): 1331-1341.
Elmanama, A. A., Al-Sheboul, S., and Abu-Dan, R. I. (2020). AntimicrobialResistance and Biofilm Formation of Pseudomonas aeruginosa. The International Arabic Journal of Antimicrobial Agents, 10(2).
Elnegery A. A., Mowafy W. K., Zahra T., El-Khier N. (2021). Study of quorum-sensing LasR and RhlR genes and their dependent virulence factors in Pseudomonas aeruginosa isolates from infected burn wounds. Access Microbiol; 3:000211.
El-Sayed Ahmed M., Zhong L. L., Shen C., Yang Y., Doi Y., Tian G. (2020). Colistin and its role in the Era of antibiotic resistance: an extended review (2000–2019). Emerging microbes and infections, 9(1), 868-885.
Farhan S. M., Ibrahim R. A., Mahran K., Hetta H., Abd El-Baky R. (2019). Antimicrobial resistance pattern and molecular genetic distribution of metallo-β-lactamases producing Pseudomonas aeruginosa isolated from hospitals in Minia, Egypt. Infect Drug Resist12:2125–2133.
Forbes B.A., Sahm D. F., Weissfeld A. S., Baron E. (2007). In Baileyand Scott's diagnostic microbiology. 2nd (ed.). Mosby, Elsevier, Inc. USA. 10840.
Ghafoor A, Jordens Z, Rehm BH. Role of PelF in pel polysaccharide biosynthesis in Pseudomonas aeruginosa. Appl Environ Microbiol. 2013;79(9):2968–2978. doi:10.1128/aem.03666-12
Hemmati F., Salehi R., Ghotaslou R., Kafil H. S., Hasani A., Gholizadeh P., Rezaee M. (2020). Quorum quenching: A potential target for antipseudomonal therapy. Infection and Drug Resistance, 13, 2989.
Hesse S. E., Adhya S. (2019). Phage Therapy in the Twenty-First Century: Facing the Decline of the Antibiotic Era; Is It Finally Time for the Age of the Phage? Annu. Rev. Microbiol, 73, 155–174.
Holbrook S. Y., Garneau-Tsodikova S. (2018). Evaluation of aminoglycoside and carbapenem resistance in a collection of drug-resistant Pseudomonas aeruginosa clinical isolates. Microbial Drug Resistance, 24(7): 1020-1030.
Hossein H. M, Mehdi R. M, Masoumeh A., Gholamreza A., Masoud D. (2015). Molecular evaluation of Pseudomonas aeruginosa isolated from patients in burn ward, ICU, and ITU, in several hospitals in Kerman province .5 (2): 1428-1431.
Impey R. E., Hawkins D. A., Sutton J., Soares da Costa T. (2020). Overcoming intrinsic and acquired resistance mechanisms associated with the cell wall of Gram-negative bacteria. Antibiotics, 9(9), 623.
Invernizzi, R., Lloyd, C. M., and Molyneaux, P. L. (2020). Respiratory microbiome and epithelial interactions shape immunity in the lungs. Immunology, 160(2), 171-182.
Jakubovics N. S., Goodman S. D., Mashburn‐Warren L., Stafford G., Cieplik F. (2021). The dental plaque biofilm matrix. Periodontology 2000, 86(1), 32-56.
Jeschke M. G.,van Baar M. E., Choudhry M., Chung K., Gibran N., Logsetty S. (2020). Burn injury Marc. Nature Reviews Disease Primers, 6(1), 1–25.
Kamalanathan M., Doyle S. M., Xu C., Achberger A., Wade T., Schwehr K., Quigg A. (2020). Exoenzymes as a signature of microbial response to marine environmental conditions. M systems, 5(2), e00290-20.
Kamali E., Jamali A., Ardebili A., Ezadi F., Mohebbi A. (2020). Evaluation of antimicrobial resistance, biofilm forming potential, and the presence of biofilm-related genes among clinical isolates of Pseudomonas aeruginosa. BMC research notes, 13, 1-6.
Lee.K., and Yoon,s.s.(2017). Pseudomonas aeruginosa biofilm, a programmed bacterial life for fitness,27(6)1053-1064.
Leiris S., Davies D. T., Sprynski N., Castandet J., Beyria L., Bodnarchuk M. S., Everett M. (2021). Virtual screening approach to identifying a novel and tractable series of Pseudomonas aeruginosa elastase inhibitors. ACS Medicinal Chemistry Letters, 12(2), 217-227.
Liao C., Huang1 X., Wang Q., Yao D., Wuyuan L. (2022). Key laboratory of medical molecular virology (MOE/ National Health Commission NHC/ CAMS), Fudan University, Shanghai, China, Shanghai.
Liao Y. C., Cheng H. W., Wu H., Kuo S., Lauderdale T., Chen F. (2019). Completing circular bacterial genomes with assembly complexity by using a sampling strategy from a single Minion run with barcoding. Frontiers in microbiology, 10, 2068-2078.
Lu M., Wang S., Wang T., Hu S., Bhayana B., Ishii M., Kong Y., Cai Y., Dai T., Cui W. (2021). Bacteria-specific phototoxic reactions triggered by blue light and phytochemical carvacrol. Sci. Transl. Med, 13, 575.
Magalhães B., Valot B., Abdelbary M., Prod'hom G., Greub G., Senn L., Blanc D. (2020). Combining standard molecular typing and whole genome sequencing to investigate Pseudomonas aeruginosa epidemiology in intensive care units. Frontiers in public health, 8(3):1-10.
Michalska M., Wolf P. (2015). Pseudomonas Exotoxin A: optimized by evolution for effective killing. Frontiers in microbiology, 6.963. Microbiol. Biotechnol. 86 (6), 1637–1645.
Minh T. T., Wibowo D., Bernd H., Rehm B. (2020). Centre for Cell Factories and Biopolymers, Griffith Institute for Drug Discovery, Griffith University, Nathan, QLD 4111, Australia.
Murray T. S., Kazmierczak B. I. (2007). Departments of Laboratory Medicine (Clinical Microbiology)1 and Medicine (Infectious Diseases) 2 and Section of Microbial Pathogenesis, 3 Yale University School of Medicine, New Haven, Connecticut 06520 p. 2700–2708 Vol. 190, No. 80021-9193.
Nathwani A. C., Reiss U., Tuddenham E., Chowdary P., McIntosh J., Riddell A., Halka K. (2018). Adeno-associated mediated gene transfer for hemophilia B: 8 years follow up and impact of removing" empty viral particles" on safety and efficacy of gene transfer.
Neha Sabharwal, Shriya Dhall, Sanjay Chhibber, Kusum Harjai (2014).Department of Microbiology, Basic Medical Sciences Block-I, South Campus, Panjab University, Chandigarh, India
Nimri L., Sulaiman M., Hani O. (2017). Community-acquired urinary tract infections caused by Burkholderia cepacia complex in patients with no underlying risk factor. JMM Case Rep;4(1): e005081.
Nitz F., de Melo B., da Silva L., Monteiro A., Marques S., Monteiro-Neto V., Turri R., et al. (2021). Molecular Detection of Drug-Resistance Genes of bla (OXA-23)-bla(OXA-51) and mcr-1 in Clinical Isolates of Pseudomonas aeruginosa. Microorganisms, 9, 786.
Obaid W. J. (2024). Molecular Detection of Biofilm Associated (RHII and rhAB) Genes in Pseudomonas aeruginosa Isolated from Different Clinical Sourses. College of Biotechnology, Al-Nahrain University, M.Sc. in Biotechnology, Supervised by Dr. Bushra Hindi Saleh, August 2024.
O’Callaghan, R., Caballero, A., Tang, A., and Bierdeman, M. (2019). Pseudomonas aeruginosa keratitis: protease IV and PASP as corneal virulence mediators. Microorganisms, 7(9), 281.
Oluwabusola E. T., Katermeran N. P., Poh W., Goh T., Tan L., Diyaolu O., Jaspars M. (2022). Inhibition of the Quorum Sensing System, Elastase Production and Biofilm Formation in Pseudomonas aeruginosa by Psammaplin A and Bisaprasin. Molecules, 27(5), 1721.
Ouyang J., Feng W., Lai X., Chen Y., Zhang X., Rong L., Chen Y. (2020). Quercetin inhibits. aeruginosa biofilm formation via the vfr-mediated lasIR system. Microbial pathogenesis, 149, 104291.
Paller A. S., Kong H. H., Seed P., Naik S., Scharschmidt T., Gallo R., Irvine A. (2019). The microbiome in patients with atopic dermatitis. Journal of Allergy and Clinical Immunology, 143(1), 26-35.
Parasuraman P., Devadatha B., Sarma V., Ranganathan S., Ampasala D., Siddhardha B. (2020). Anti-quorum sensing and antibiofilm activities of Blastobotrys parvus PPR3 against Pseudomonas aeruginosa PAO1. Microbial pathogenesis, 138, 103811.
Parsa, P., Amirmozafari, N., Nowruzi, B., & Bahar, M. A. (2020). Molecular characterization of polymorphisms among Pseudomonas aeruginosa strains isolated from burn patients' wounds. Heliyon, 6(12), e05041
Phan J., Ranjbar S., Kagawa M., Gargus M., Hochbaum A., Whiteson K. (2020). Thriving under stress: Pseudomonas aeruginosa outcompetes the background polymicrobial community under treatment conditions in a novel chronic wound model. Frontiers in Cellular and Infection Microbiology. Jundishapur, J. Microbiol. e90034: 1-8.
Rendueles O., Ghigo J.-M. (2012). Multi-species biofilms: how to avoid unfriendly neighbors. FEMS Microbiol. Rev. 36, 972–989.
Riquelme S. A., Liimatta K., Lung T., Fields B., Ahn D., Chen D., Prince A. (2020). Pseudomonas aeruginosa utilize host-derived itaconate to redirect its metabolism to promote biofilm formation. Cell metabolism, 31(6), 1091-1106.
Saleem S., Bokhari H. (2020). Journal of Infection and Public Health 13 598–605 Department of Biosciences, Comsats University, Islamabad, Pakistan
Schreiber K., Krieger R., Benkert B., Eschbach M., Arai H., Schobert M., Jahn D. (2007). The anaerobic regulatory networkrequired for Pseudomonas aeruginosa nitrate respiration. J Bacteriol189, 4310–4314.
Stones, D.H. and Krachler, A. (2015). Fatal attraction: how bacterial adhesins affect host signaling and what we can learn from them. Int. J.Mol. Sci. 16: 2626-2640.
Shaaban S., Al-Qahtani A., Mohammed Al-Ahdal M., Barwa R. (2017). Molecular characterization of resistance mechanisms in Pseudomonas aeruginosa isolates resistant to carbapenems. J. Infect. Dev. Ctries. 11(12): 935-943.
Shariati A., Asadian E., Fallah F., Azimi T., Hashemi A., Yasbolaghi Sh. J., Taati M. M. (2019). Evaluation of nano curcumineffects on expression levels of virulence genes and biofilm production of multidrug-resistant Pseudomonas aeruginosa isolated from burn wound infection in Tehran, Iran. Infection and drug resistance, 2223- 2235.
Shineh G., Mobaraki M., Bappy M. J., K. Mills D. (2023). Biofilm formation, and related impacts on healthcare, food processing and packaging, industrial manufacturing, marine industries, and sanitation–A review. Applied Microbiology 3, 629–665.
Swapna M., Sumathi G., Anitha M. (2022). Correlation of biofilm production with antibiotic susceptibility pattern of Pseudomonas aeruginosa from various clinical specimens. Asian Journal of Medical Sciences, 13(1), 88-92.
Tacconelli, E., Carrara, E., Savoldi, A., Harbarth, S., Mendelson, M., Monnet, D. L., and Zorzet, A. (2018). Discovery, research, and development of new antibiotics: the WHO priority list of antibioticresistant bacteria and tuberculosis. The Lancet Infectious Diseases, 18(3), 318-327
Webster S. S., Lee C. K., Schmidt W. C., Wong G., O'Toole G. (2021). Interaction between the type 4 pili machinery and a diguanylate cyclase fine-tune C-Di-GMP levels during early biofilm formation. Proc. Natl. Acad. Sci. United States America 118 (26), e2105566118.
Welde N., Abunna F., Wodajnew B. (2020). Isolation, identification and antimicrobial susceptibility profiles of E. coliO157: H7 from raw cow milk in and aroundModjotown, Ethiopia. J. American Science. 16(6).
Yang F., Liu C., Ji J., Cao W., Ding B., Xu X. (2021). Molecular characteristics, antimicrobial resistance, and biofilm formation of Pseudomonas aeruginosa isolated from patients with aural infections in Shanghai, China. Infection and Drug Resistance, 14, 3637.
Zadeh R. G., Kalani B. S., Ari M., Talebi M., Razavi S., Jazi F. (2022). Isolation of persister cells within the biofilm and relative gene expression analysis of type II toxin/antitoxin system in Pseudomonas aeruginosa isolates in exponential and stationary phases. Journal of global antimicrobial resistance, 28, 30-37
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