Biofilm formation and antimicrobial resistance in Enterobacterales isolated from cases of septicemia-Protocol

Biofilm formation and antimicrobial resistance in Enterobacterales isolated from cases of septicemia-Protocol | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Study protocol Biofilm formation and antimicrobial resistance in Enterobacterales isolated from cases of septicemia-Protocol Dr. Akoijam Nisha Devi, Dr. Gargi Mudey This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4407412/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Introduction: This is a presentation of a study protocol in order to evaluate biofilm formation in enterobacterales causing septicemia so that clinical issues regarding increase in infections due to antimicrobial resistance can be solved. Materials and methods: This is a study protocol of a prospective cross sectional simple random sampling method. A total of 100 patients blood samples will be included in the study. The bacteria from blood culture bottles will be isolated and identified using Bac-T Alert 3D/VITEK ID/Conventional method. Biofilm formation from cases of septicemia will be identified by tube method and tissue culture plate method. Presence of biofilm production will be detected by tube method .and tissue culture plate method Reference strain of positive biofilm producer Klebsiella pneumoniae 700603 will be used as control. Isolates positive by both the methods will be labelled biofilm producers. Their antimicrobial resistance will be studied. Antibiotic susceptibility will be carried out by Kirby Bauer disc diffusion method as per CLSI recommendations. The medium, Muller Hilton Agar (MHA) will be used. Quality control strains of Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), and Pseudomonas aeruginosa (ATCC 27853) will be used to validate the results of antibiotic susceptibility test. Results: This is an ongoing study protocol, and we are expecting the analysis of the results in July 2024. Conclusion: The ability of Enterobacterales to form biofilms and acquire antimicrobial resistance significantly contributes to the pathogenesis of septicemia and treatment failure. Understanding the mechanism of formation and antimicrobial resistance will help design effective treatment and therapeutic strategies. Categories: Microbiology, experiment septicemia Enterobacterales biofilm formation antimicrobial resistance treatment failure INTRODUCTION Septicemia is a condition in which bacteria circulate and actively multiply in the bloodstream and produce toxins, resulting in overwhelming host immune response that cause harm to the host[ 1 ]. Bacteria belonging to Enterobacterales are among the most common organisms causing septicemia[ 2 ]. Biofilms are microbial-derived surface-associated cells that are enclosed in an extracellular polymeric substance matrix and attached to substratum or to each other in an irreversible way. Biofilms are important virulence factors which are produced through a serial process beginning with a single species of bacteria with fimbria, pili or flagella attaching to the conditioning film then progressing to form microcolonies and colonies after longer exposure eventually forms a mass that detaches after maturation 3 . Bacteria form biofilm as a part of their survival mechanisms[ 4 ]. Biofilm formation is a serious clinical issue because biofilms promotes antimicrobial resistance by slowing down antimicrobial diffusion and also by facilitating plasmid exchange[ 3 ].Biofilm formation is also associated with the severity, persistence and relapse of infection[ 3 ] For Enterobacterales resistance to the beta lactam class of antibiotics is particularly concerning because they are the cornerstone of treatment for these infections. Resistance to beta lactam antibiotics can occur via efflux pumps, altered penicillin binding sites and beta lactamases which cleave the beta lactam ring and inactivate the target antibiotic[ 2 ] The present study will be conducted to find out the bacteria belonging to Enterobacterales family causing septicaemia in our community and their ability to form biofilms and also to correlate the antimicrobial resistance pattern of the organisms with biofilm forming capacity. The present study will guide the clinicians to use proper antimicrobials in patients with septicaemia in our community in the geographical region in and around the hospital. MATERIALS AND METHODS The study will be performed in Department of Microbiology, Jawaharlal Nehru Medical College Sawangi(Meghe) Wardha. Funding declaration : It is funded by Datta Meghe Institute of Higher Education and Research. Approval from the Institutional Ethical Committee has been taken (DMIMS(DU)/IEC/748/2023). Protocol is registered on clinical trials REF/2024/01/077896. Study design: A total of 100 blood culture bottles from patients will be obtained. It will follow prospective cross sectional study design and simple random sampling is done for these samples. Methods : Positive blood culture bottles will be detected by Bac-T Alert 3D. The organism will be isolated by subculturing it in Blood agar, MacConkey and gram staining methods. It can be identified by VITEK ID/ conventional method. Biofilm formation from cases of septicemia will be identified by tube method and tissue culture plate method[5]. Presence of biofilm production will be detected by tube method .and tissue culture plate method. Reference strain of positive biofilm producer Klebsiella pneumoniae 700603 will be used as control. Isolates positive by both the methods will be labelled biofilm producers. Their antimicrobial resistance will be studied. Antibiotic susceptibility will be carried out by Kirby Bauer disc diffusion method as per CLSI recommendations[6]. The medium, Muller Hilton Agar (MHA) will be used. Quality control strains of Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), and Pseudomonas aeruginosa (ATCC 27853) will be used to validate the results of antibiotic susceptibility test. Association between antimicrobial resistance patterns and biofilm formation will also be analyzed. Inclusion criteria : Patients whose blood cultures are positive by Bac-TAlert-3D and Enterobacterales isolated from their blood cultures by conventional/Vitek method are included in the study. Exclusion criteria : Gram positive organisms, contaminants, candida and bacteria not belonging to Enterobacterales isolated from the patients are excluded from the study. Data collection methods : WHONET software is used to collect data. Version 2022. WHONET is a free desktop Windows application for the management and analysis of microbiology laboratory data with a particular focus on antimicrobial resistance surveillance developed and supported by the WHO collaborating center for surveillance of antimicrobial resistance at the Brigham and Women’s Hospital in Boston, Massachusetts. It can provide guidance for empiric therapy of infections, alert clinicians of trends of antimicrobial resistance, guide drug policy decisions and preventive measures. Outcome measures: Number of organism isolated, type of organisms isolated, Number of isolates producing biofilm and association between antibiotic resistance and biofilm formation. Sample size: 100 non-repeat enterobacterales isolated from blood cultures of clinical diagnosed septicaemia cases from Jan 2023 to Dec 2024 will be included in the study. Statistical Methods : Data so collected will be entered in the worksheet of SPSS Software version (16.0) and chi square test will be performed to find the association between biofilm forming capacity and resistance to an antibiotic of an organism. Chi square test will also be performed to find the association between biofilm forming capacity and multi drug resistance status of an organism. P value be calculated and considered significant only when it will be less than or equal to 0.05. RESULTS This is an ongoing study protocol, and we are expecting the analysis of the results in July 2024. DISCUSSION Bacteria growing in a biofilm are sessile and are responsible for most physiologic processes in the biofilm environment[ 7 ]. They are responsible for approximately 70% of all human microbial infections[ 8 ]. Enterobacterales are frequently found in a biofilm. Biofilm formation involves several stages such as attachment, irreversible adhesion, microcolony formation, growth and maturation, dispersion[ 8 ]. Biofilm is the natural mode of bacterial growth in nature. Being a survival strategy of bacteria, biofilms may grow in the primary focus of infection and provide dissemination of bacteria by the blood stream. Another source of bacterial biofilm in the bloodstream is indwelling blood contacting medical devices (arterial and venous catheters and prostheses, mechanical heart valves, etc.). Enterobacterales are a major group of organisms found in the biofilm formed on these medical devices. Initially composed of single species, but longer exposure inevitably leads to multispecies biofilms[ 9 ].Considering, all these it is obvious that, biofilm plays a crucial role in the pathogenesis of septicemia[ 4 ]. Microbial biofilm is the most defensive strategy adopted by bacteria. Biofilms protect the microbial community from external damage. Bacterial biofilms are highly resistant to antibiotic treatment and immune responses. In comparison with planktonic bacterial cultures, biofilm formation leads to a large increase (upto 1000fold) in resistance to antimicrobial agents. The adequate concentration of antibiotic for eradication of mature biofilm is difficult to reach invivo[ 10 ]. The Enterobacterales like E.coli and Klebsiella pneumoniae are among the common organisms which cause septicemia[ 14 ]. Recently there has been a vicious cycle of increasing antimicrobial resistance among the Enterobacterales, specially with E. coli and K. pneumoniae. Worldwide dissemination of extended spectrum beta lactamases(ESBLs) among Enterobacterales has resulted in increased resistance to all penicillin and cephalosporin antibiotics. This has led to an increase in the use of carbapenem. But the increased use of carbapenem has led to the birth and spread of carbapenem resistant Enterobacterales all over the world[ 11 , 12 , 13 ]. Infections caused by CRE are a major health problem due to the limited therapeutic options and high mortality rates, associated with these infections. There are three major classes of carbapenem resistant (CR) determinants. They are classified as Class A, Klebsiella pneumoniae carbapenemase (KPC), Class B, Metallo-ß-Lactamases(MBLs), such as New Delhi MBL (NDM), Verona integrin-encoded MBL (VIM) and imipenemase(IMP) and Class D, oxacillinases(OXA) -type enzymes such as OXA-48 like carbapenemases[ 15 , 16 ]. Biofilm formation poses a significant challenge in clinical settings, as it leads to antimicrobial resistance in Enterobacterales leading to treatment failures and the persistence of infections. CONCLUSION The ability of Enterobacterales to form biofilms and acquire antimicrobial resistance significantly contributes to the pathogenesis of septicemia and treatment failure. Understanding the mechanisms of biofilm formation and the development of antimicrobial resistance is crucial for the design of effective therapeutic strategies. Future research efforts should focus on developing novel antimicrobial agents and innovative approaches to disrupt biofilms, ultimately improving the management and outcome of septicemia caused by Enterobacterales. Declarations Author Contribution Dr. Akoijam Nisha wrote the main manuscript text, performed all the experiments, designed and implemented the study and Dr. Gargi Mudey guided in the study design, statistical analysis and implementation. Data Availability Data is link to this webpage http://go.microsoft.com/fwlink/p/?LinkId=255141 References Tille PM. Bailey and Scott’s Diagnostic Microbiology. Ed 14, St. Louis Missouri. Elsevier; 2017. Tompkins K, Juliano JJ, van Duin D. Antimicrobial Resistance in Enterobacterales and Its Contribution to Sepsis in Sub-saharan Africa. Front Med (Lausanne). 2021;8:615649. 10.3389/fmed.2021.615649 . PMID: 33575265; PMCID: PMC7870712. Juliana A, Leela KV, Gopinathan A, Jayaprakash T. Biofilm Formation and its Association with Gram Negative Sepsis Pathogenicity. Biomedical Pharmacol J. 2022;15(4):2099–106. Vestby LK, Grønseth T, Simm R, Nesse LL. Bacterial Biofilm and its Role in the Pathogenesis of Disease. Antibiot (Basel). 2020;9(2):59. 10.3390/antibiotics9020059 . PMID: 32028684; PMCID: PMC7167820. Christensen GD, Simpson WA, Younger JJ, Baddour LM, Barrett FF, Melton DM, Beachey EH. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol. 1985;22(6):996–1006. 10.1128/jcm.22.6.996-1006.1985 . PMID: 3905855; PMCID: PMC271866. Clinical and laboratory standards. Institute performance standards for antimicrobial susceptibility testing; Thirty second edition CLSI supplement M100. Clinical and Laboratory Standards Institute; 2022. Berlanga M, Guerrero R. Living together in biofilms: the microbial cell factory and its biotechnological implications. Microb Cell Fact. 2016;15(1):165. 10.1186/s12934-016-0569-5 . PMID: 27716327; PMCID: PMC5045575. .Schulze A, Mitterer F, Pombo JP, Schild S. Biofilms by bacterial human pathogens: Clinical relevance - development, composition and regulation - therapeutical strategies. Microb Cell. 2021;8(2):28–56. 10.15698/mic2021.02.741 . PMID: 33553418; PMCID: PMC7841849. Hernández-Jiménez E, Del Campo R. Toledano Biofilm vs. planktonic bacterial mode of growth: which do human macrophages prefer? Biochem Biophys Res Commun. 2013;441(4):947–52. Epub 2013 Nov 14. PMID: 24239884. de la Fuente-Núñez C, Reffuveille F, Fernández L, Hancock RE. Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies. Curr Opin Microbiol. 2013;16(5):580–9. 10.1016/j.mib.2013.06.013 . Epub 2013 Jul 20. PMID: 23880136. Meyer E, Schwab F, Schroeren-Boersch B, Gastmeier P. Dramatic increase of third-generation cephalosporin-resistant E. coli in German intensive care units: secular trends in antibiotic drug use and bacterial resistance, 2001 to 2008. Crit Care. 2010;14(3):R113. 10.1186/cc9062 . Epub 2010 Jun 14. PMID: 20546564; PMCID: PMC2911759. Codjoe FS, Donkor ES. Carbapenem Resistance: A Review. Med Sci (Basel). 2017;6(1):1. 10.3390/medsci6010001 . PMID: 29267233; PMCID: PMC5872158. Lutgring JD. Carbapenem-resistant Enterobacteriaceae: An emerging bacterial threat. Semin Diagn Pathol. 2019;36(3):182–6. 10.1053/j.semdp.2019.04.011 . Epub 2019 Apr 17. PMID. Ajulo S, Awosile B. Global antimicrobial resistance and use surveillance system (GLASS 2022): Investigating the relationship between antimicrobial resistance and antimicrobial consumption data across the participating countries. PLoS ONE. 2024;19(2):e0297921. 10.1371/journal.pone.0297921 . PMID: 38315668; PMCID: PMC10843100. Magiorakos AP, Burns K, Rodríguez Baño J, Borg M. Infection prevention and control measures and tools for the prevention of entry of carbapenem-resistant Enterobacteriaceae into healthcare settings: guidance from the EuropeanCentre for Disease Prevention and Control. Antimicrob Resist Infect Control. 2017;6:113. 10.1186/s13756-017-0259-z . PMID: 29163939; PMCID: PMC5686856. Nordmann P, Dortet L, Poirel L. Carbapenem resistance in Enterobacteriaceae: here is the storm! Trends Mol Med. 2012;18(5):263 – 72. 10.1016/j.molmed.2012.03.003 . Epub 2012 Apr 3. PMID: 22480775. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4407412","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Study protocol","associatedPublications":[],"authors":[{"id":305198590,"identity":"46365c06-402a-46b3-a211-91d261ede118","order_by":0,"name":"Dr. Akoijam Nisha Devi","email":"","orcid":"","institution":"JNMC,DMIHER sawangi wardha maharashtra","correspondingAuthor":false,"prefix":"Dr.","firstName":"Akoijam","middleName":"Nisha","lastName":"Devi","suffix":""},{"id":305198591,"identity":"8b258433-efd5-4ca0-9039-7de8b326b0e6","order_by":1,"name":"Dr. Gargi Mudey","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9ElEQVRIiWNgGAWjYLCCBAYGxgYG5mNgDhs78VrY0sAsNmYiLQJq4TEDa2EgpEXe/ezjFw9zDsv2S+R8e/DxxzZ5PmYGxg8fc3BrMTyTbmaRuO2w8cwZudsNZyTcNmxjZmCWnLkNj5aGNDYDoJbEDTdyt0nzJNxmBGphY+bFp6X/GUTL/hs5z0Ba7AlqkZdIY34AtkUihw2kJZGgFgOJZ2wMidvSjWeceWYmOSPtdnIbM2MzXr/I96cxf/y5zVq2vz35mcQHm9u289ubD374iM+WAwxsEmCWQAJMDBhH+IA8MKF8ALP4D+BVOApGwSgYBSMYAACMJ1NqXySMNQAAAABJRU5ErkJggg==","orcid":"","institution":"JNMC,DMIHER sawangi wardha maharashtra","correspondingAuthor":true,"prefix":"Dr.","firstName":"Gargi","middleName":"","lastName":"Mudey","suffix":""}],"badges":[],"createdAt":"2024-05-12 06:23:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4407412/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4407412/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":64082262,"identity":"35762ec9-9c0c-4f07-91ae-2c09e3e1254a","added_by":"auto","created_at":"2024-09-06 10:45:12","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":247898,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4407412/v1/fdcdd7e2-b719-44f3-90e1-d2c5b66d0f58.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Biofilm formation and antimicrobial resistance in Enterobacterales isolated from cases of septicemia-Protocol","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eSepticemia is a condition in which bacteria circulate and actively multiply in the bloodstream and produce toxins, resulting in overwhelming host immune response that cause harm to the host[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Bacteria belonging to Enterobacterales are among the most common organisms causing septicemia[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBiofilms are microbial-derived surface-associated cells that are enclosed in an extracellular polymeric substance matrix and attached to substratum or to each other in an irreversible way. Biofilms are important virulence factors which are produced through a serial process beginning with a single species of bacteria with fimbria, pili or flagella attaching to the conditioning film then progressing to form microcolonies and colonies after longer exposure eventually forms a mass that detaches after maturation\u003csup\u003e3\u003c/sup\u003e. Bacteria form biofilm as a part of their survival mechanisms[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Biofilm formation is a serious clinical issue because biofilms promotes antimicrobial resistance by slowing down antimicrobial diffusion and also by facilitating plasmid exchange[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].Biofilm formation is also associated with the severity, persistence and relapse of infection[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]\u003c/p\u003e \u003cp\u003eFor Enterobacterales resistance to the beta lactam class of antibiotics is particularly concerning because they are the cornerstone of treatment for these infections. Resistance to beta lactam antibiotics can occur via efflux pumps, altered penicillin binding sites and beta lactamases which cleave the beta lactam ring and inactivate the target antibiotic[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/p\u003e \u003cp\u003eThe present study will be conducted to find out the bacteria belonging to Enterobacterales family causing septicaemia in our community and their ability to form biofilms and also to correlate the antimicrobial resistance pattern of the organisms with biofilm forming capacity. The present study will guide the clinicians to use proper antimicrobials in patients with septicaemia in our community in the geographical region in and around the hospital.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eThe study will be performed in Department of Microbiology, Jawaharlal Nehru Medical College Sawangi(Meghe) Wardha.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding declaration\u003c/strong\u003e: It is funded by Datta Meghe Institute of Higher Education and Research. Approval from the Institutional Ethical Committee has been taken (DMIMS(DU)/IEC/748/2023).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eProtocol is registered on clinical trials REF/2024/01/077896.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStudy design:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA total of 100 blood culture bottles from patients will be obtained. It will follow prospective cross sectional study design and simple random sampling is done for these samples.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: Positive blood culture bottles will be detected by Bac-T Alert 3D. The organism will be isolated by subculturing it in Blood agar, MacConkey and gram staining methods. It can be identified by VITEK ID/ conventional method.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBiofilm formation from cases of septicemia will be identified by tube method and tissue culture plate method[5].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePresence of biofilm production will be detected by tube method .and tissue culture plate\u0026nbsp;\u003c/p\u003e\n\u003cp\u003emethod. Reference strain of positive biofilm producer Klebsiella pneumoniae 700603 will be\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eused as control. Isolates positive by both the methods will be labelled biofilm producers.\u003c/p\u003e\n\u003cp\u003eTheir antimicrobial resistance will be studied.\u003c/p\u003e\n\u003cp\u003eAntibiotic susceptibility will be carried out by Kirby Bauer disc diffusion method as per\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCLSI recommendations[6]. The medium, Muller Hilton Agar (MHA) will be used.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eQuality control strains of Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e25923), and Pseudomonas aeruginosa (ATCC 27853) will be used to validate the results of antibiotic susceptibility test.\u003c/p\u003e\n\u003cp\u003eAssociation between antimicrobial resistance patterns and biofilm formation will also be analyzed.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInclusion criteria\u003c/strong\u003e: Patients whose blood cultures are positive by Bac-TAlert-3D and Enterobacterales isolated from their blood cultures by conventional/Vitek method are included in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExclusion criteria\u003c/strong\u003e: Gram positive organisms, contaminants, candida and bacteria not belonging to Enterobacterales isolated from the patients are excluded from the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData collection methods\u003c/strong\u003e:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWHONET software is used to collect data. Version 2022.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; WHONET is a free desktop Windows application for the management and analysis of\u0026nbsp;\u003c/p\u003e\n\u003cp\u003emicrobiology laboratory data with a particular focus on antimicrobial resistance surveillance\u0026nbsp;\u003c/p\u003e\n\u003cp\u003edeveloped and supported by the WHO collaborating center for surveillance of antimicrobial\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eresistance at the Brigham and Women’s Hospital in Boston, Massachusetts.\u003c/p\u003e\n\u003cp\u003eIt can provide guidance for empiric therapy of infections, alert clinicians of trends of\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eantimicrobial resistance, guide drug policy decisions and preventive measures.\u003c/p\u003e\n\u003cp\u003eOutcome measures:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eNumber of organism isolated, type of organisms isolated, Number of isolates producing biofilm and association between antibiotic resistance and biofilm formation.\u003c/p\u003e\n\u003cp\u003eSample size:\u003c/p\u003e\n\u003cp\u003e100 non-repeat enterobacterales isolated from blood cultures of clinical diagnosed septicaemia cases from Jan 2023 to Dec 2024 will be included in the study.\u003c/p\u003e\n\u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical Methods\u003c/strong\u003e:\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;Data so collected will be entered in the worksheet of SPSS Software version (16.0) and chi square test will be performed to find the association between biofilm forming capacity and resistance to an antibiotic of an organism. Chi square test will also be performed to find the association between biofilm forming capacity and multi drug resistance status of an organism. P value be calculated and considered significant only when it will be less than or equal to 0.05.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003eThis is an ongoing study protocol, and we are expecting the analysis of the results in July 2024.\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eBacteria growing in a biofilm are sessile and are responsible for most physiologic processes in the biofilm environment[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. They are responsible for approximately 70% of all human microbial infections[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Enterobacterales are frequently found in a biofilm.\u003c/p\u003e \u003cp\u003eBiofilm formation involves several stages such as attachment, irreversible adhesion, microcolony formation, growth and maturation, dispersion[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Biofilm is the natural mode of bacterial growth in nature. Being a survival strategy of bacteria, biofilms may grow in the primary focus of infection and provide dissemination of bacteria by the blood stream.\u003c/p\u003e \u003cp\u003eAnother source of bacterial biofilm in the bloodstream is indwelling blood contacting\u003c/p\u003e \u003cp\u003emedical devices (arterial and venous catheters and prostheses, mechanical heart valves, etc.).\u003c/p\u003e \u003cp\u003eEnterobacterales are a major group of organisms found in the biofilm formed on these medical devices. Initially composed of single species, but longer exposure inevitably leads to multispecies biofilms[\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].Considering, all these it is obvious that, biofilm plays a crucial role in the pathogenesis of septicemia[\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMicrobial biofilm is the most defensive strategy adopted by bacteria. Biofilms protect the microbial community from external damage. Bacterial biofilms are highly resistant to antibiotic treatment and immune responses. In comparison with planktonic bacterial cultures, biofilm formation leads to a large increase (upto 1000fold) in resistance to antimicrobial agents. The adequate concentration of antibiotic for eradication of mature biofilm is difficult to reach invivo[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe Enterobacterales like E.coli and Klebsiella pneumoniae are among the common organisms which cause septicemia[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Recently there has been a vicious cycle of increasing antimicrobial resistance among the Enterobacterales, specially with E. coli and K. pneumoniae. Worldwide dissemination of extended spectrum beta lactamases(ESBLs) among Enterobacterales has resulted in increased resistance to all penicillin and cephalosporin antibiotics. This has led to an increase in the use of carbapenem. But the increased use of carbapenem has led to the birth and spread of carbapenem resistant Enterobacterales all over the world[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eInfections caused by CRE are a major health problem due to the limited therapeutic options and high mortality rates, associated with these infections. There are three major classes of carbapenem resistant (CR) determinants. They are classified as Class A, Klebsiella pneumoniae carbapenemase (KPC), Class B, Metallo-\u0026szlig;-Lactamases(MBLs), such as New Delhi MBL (NDM), Verona integrin-encoded MBL (VIM) and imipenemase(IMP) and Class D, oxacillinases(OXA) -type enzymes such as OXA-48 like carbapenemases[\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBiofilm formation poses a significant challenge in clinical settings, as it leads to antimicrobial\u003c/p\u003e \u003cp\u003eresistance in Enterobacterales leading to treatment failures and the persistence of infections.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThe ability of Enterobacterales to form biofilms and acquire antimicrobial resistance significantly contributes to the pathogenesis of septicemia and treatment failure. Understanding the mechanisms of biofilm formation and the development of antimicrobial resistance is crucial for the design of effective therapeutic strategies. Future research efforts should focus on developing novel antimicrobial agents and innovative approaches to disrupt biofilms, ultimately improving the management and outcome of septicemia caused by Enterobacterales.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eDr. Akoijam Nisha wrote the main manuscript text, performed all the experiments, designed and implemented the study and Dr. Gargi Mudey guided in the study design, statistical analysis and implementation.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eData is link to this webpage http://go.microsoft.com/fwlink/p/?LinkId=255141\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eTille PM. 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Trends Mol Med. 2012;18(5):263\u0026thinsp;\u0026ndash;\u0026thinsp;72. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.molmed.2012.03.003\u003c/span\u003e\u003cspan address=\"10.1016/j.molmed.2012.03.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2012 Apr 3. PMID: 22480775.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"septicemia, Enterobacterales, biofilm formation, antimicrobial resistance, treatment failure","lastPublishedDoi":"10.21203/rs.3.rs-4407412/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4407412/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIntroduction: This is a presentation of a study protocol in order to evaluate biofilm formation in enterobacterales causing septicemia so that clinical issues regarding increase in infections due to antimicrobial resistance can be solved.\u003c/p\u003e\n\u003cp\u003eMaterials and methods: This is a study protocol of a prospective cross sectional simple random sampling method. A total of 100 patients blood samples will be included in the study. The bacteria from blood culture bottles will be isolated and identified using Bac-T Alert 3D/VITEK ID/Conventional method. Biofilm formation from cases of septicemia will be identified by tube method and tissue culture plate method.\u003c/p\u003e\n\u003cp\u003ePresence of biofilm production will be detected by tube method .and tissue culture plate\u003c/p\u003e\n\u003cp\u003emethod Reference strain of positive biofilm producer Klebsiella pneumoniae 700603 will be\u003c/p\u003e\n\u003cp\u003eused as control. Isolates positive by both the methods will be labelled biofilm producers.\u003c/p\u003e\n\u003cp\u003eTheir antimicrobial resistance will be studied.\u003c/p\u003e\n\u003cp\u003eAntibiotic susceptibility will be carried out by Kirby Bauer disc diffusion method as per\u003c/p\u003e\n\u003cp\u003eCLSI recommendations. The medium, Muller Hilton Agar (MHA) will be used.\u003c/p\u003e\n\u003cp\u003eQuality control strains of Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC\u003c/p\u003e\n\u003cp\u003e25923), and Pseudomonas aeruginosa (ATCC 27853) will be used to validate the results of antibiotic susceptibility test.\u003c/p\u003e\n\u003cp\u003eResults: This is an ongoing study protocol, and we are expecting the analysis of the results in July 2024.\u003c/p\u003e\n\u003cp\u003eConclusion: The ability of Enterobacterales to form biofilms and acquire antimicrobial resistance significantly contributes to the pathogenesis of septicemia and treatment failure. Understanding the mechanism of formation and antimicrobial resistance will help design effective treatment and therapeutic strategies.\u003c/p\u003e\n\u003cp\u003eCategories: Microbiology, experiment\u003c/p\u003e","manuscriptTitle":"Biofilm formation and antimicrobial resistance in Enterobacterales isolated from cases of septicemia-Protocol","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-05-24 19:35:06","doi":"10.21203/rs.3.rs-4407412/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f3ed7cca-ea8f-41ad-b48e-a050676c06d8","owner":[],"postedDate":"May 24th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-09-06T10:37:06+00:00","versionOfRecord":[],"versionCreatedAt":"2024-05-24 19:35:06","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4407412","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4407412","identity":"rs-4407412","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}