en Anti-microbial agents, resistance and treatment protocols Anti-microbial agents, resistance and treatment protocols Original article Original article <b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Introduction: </span></span></span></b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Biofilms are often found in communities of microorganisms in chronic and persistent infections, exhibiting high resistance against antimicrobial agents. Biofilm serves as a barrier, impeding the penetration of drugs and constraining their effectiveness. Multiple methods, such as the Tissue Culture Plate method, Congo Red Agar method, Tube method, bioluminescent assay, and fluorescent microscopic examination, can be used to evaluate biofilm production</span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">. <b><span style="letter-spacing:-.05pt">Methods</span></b><span style="letter-spacing:-.05pt">:</span> The study included a total of 300 clinical isolates representing a range of bacterial species, including <i>Acinetobacter baumannii</i> (n=9), Coagulase Negative Staphylococcus (n=7), <i>Enterobacter aerogenes</i> (n=7), <i>Enterococcus faecalis</i> (n=15), <i>Escherichia coli</i> (n=137), <i>Klebsiella pneumoniae</i> (n=23), <i>Proteus mirabilis</i> (n=4), <i>Pseudomonas aeruginosa</i> (n=16), <i>Salmonella typhi</i> (n=11), and <i>Staphylococcus aureus</i> (n=68). Associations among isolates capable and incapable of biofilm formation and their multidrug resistance phenotypes were evaluated. <b>Results</b>: Among the 300 clinical isolates tested, 289 isolates (96.3%) exhibited biofilm formation.<span style="letter-spacing:.05pt"> The most prevalent biofilm-forming organisms were <i>A. baumannii</i> (n=9), <i>Citrobacter koseri</i> (n=1), Coagulase Negative Staphylococcus (CONS) (n=7), <i>E. aerogenes</i> (n=7), <i>E. faecalis</i> (n=15), <i>E. coli</i> (n=137), <i>Klebsiella oxytoca</i> (n=1), <i>K. pneumoniae</i> (n=23), <i>P. mirabilis</i> (n=4), <i>P. aeruginosa</i> (n=16), <i>S. typhi</i> (n=11), <i>S. aureus</i> (n=68), and <i>Streptococcus pneumoniae</i> (n=1). </span>The biofilm-forming isolates demonstrated increased resistance compared to isolates that did not form biofilms. <b>Conclusion</b>:<span style="letter-spacing:.05pt"> Antimicrobial resistance represents a critical characteristic of infections involving biofilms</span>. The study identified biofilm production in 92.7% of the isolates tested via TCP and in 72.3% of the isolates using the CRA. Furthermore, it was observed that pathogens with multidrug resistance (MDR) exhibited a higher biofilm production tendency than non-producing pathogens.</span></span></span><br> &nbsp; Biofilm, Resistance, Pathogens, Chronic infection, Multidrug resistance, Antimicrobial agents 148 154 http://jommid.pasteur.ac.ir/browse.php?a_code=A-10-434-1&slc_lang=en&sid=1 Rubina Shrestha 0000-0003-3930-6990 No Department of Microbiology, Modern Technical College, Pokhara University, Kathmandu, Nepal Pooja Ghaju 0000-0002-1346-7908 No Department of Microbiology, Modern Technical College, Pokhara University, Kathmandu, Nepal Dilip Kumar Chaudhary 0000-0002-1087-5156 No Department of Microbiology, Modern Technical College, Pokhara University, Kathmandu, Nepal Raushan Kumar Karn 0000-0002-5104-5597 No Department of Microbiology, Modern Technical College, Pokhara University, Kathmandu, Nepal Rajesh Kumar Thakur No Department of Microbiology, Modern Technical College, Pokhara University, Kathmandu, Nepal Suresh Jaiswal 0000-0001-5684-6994 No School of Health and Allied Sciences, Pokhara University, Pokhara, Nepal Ram Krishna Shrestha rknasika008@gmail.com 0000-0002-0193-3000 Yes Department of Microbiology, Modern Technical College, Pokhara University, Kathmandu, Nepal